bims-enlima 2025-02-09 papers

bims-enlima

Biomed News

on Engineered living materials

Issue of 2025–02–09
thirty papers selected by
Rahul Kumar, Tallinna Tehnikaülikool

Design and regulation of engineered bacteria for environmental release.
Synthetic transmembrane DNA receptors enable engineered sensing and actuation.
Engineered microbial living matter for diagnostics, prevention, and therapy.
A thiol-ene click-based strategy to customize injectable polymer-nanoparticle hydrogel properties for therapeutic delivery.
Covalent Dynamic DNA Networks to Translate Multiple Inputs into Programmable Outputs.
From 3D to 4D printing of lignin towards green materials and sustainable manufacturing.
Matrix Stiffness Regulates GBM Migration and Chemoradiotherapy Responses via Chromatin Condensation in 3D Viscoelastic Matrices.
Spatially Controlled Self-Assembly of Supramolecular Hydrogels Enabled by Light-Triggered Catalysis.
A new flavor of synthetic yeast communities sees the light.
H-bonded organic frameworks as ultrasound-programmable delivery platform.
3D-Printed Protein-Based Bioplastics with Tunable Mechanical Properties Using Glycerol or Hyperbranched Poly(glycerol)s as Plasticizers.
Engineering a genomically recoded organism with one stop codon.
Rapid isolation and recovery of Salmonella using hollow glass microspheres coated with multilayered nanofilms.
Harnessing photosynthesis for materials, devices, and environmental technologies.
Immobilization of BMP-2 in porous hydrogels to spatially regulate osteogenesis.
Bioprinting spatially guided functional 3D neural circuits with agarose-xanthan gum copolymer hydrogels.
Biological Polymers: Evolution, Function, and Significance.
Surface functionalization of microscaffolds produced by high-resolution 3D printing: A new layer of freedom.
Proteins for Hyperelastic Materials.
Synthetic Dual-Input Hybrid Riboswitches─Optimized Genetic Regulators in Yeast.
Engineered Living Systems Based on Gelatin: Design, Manufacturing, and Applications.
DRIMS: A Synthetic Biology Platform that Enables Deletion, Replacement, Insertion, Mutagenesis, and Synthesis of DNA.
Engineering microalgal cell wall-anchored proteins using GP1 PPSPX motifs and releasing with intein-mediated fusion.
Spatial propagation of temperate phages within and among biofilms.
Programming biological communication between distinct membraneless compartments.
Crosslink strength governs yielding behavior in dynamically crosslinked hydrogels.
New trends of additive manufacturing using materials based-on natural fibers and minerals : A systematic review.
A polysaccharide-based hydrogel platform for tumor spheroid production and anticancer drug screening.
Polysulfides promote protein disulfide bond formation in microorganisms growing under anaerobic conditions.
Optogenetic engineering for ion channel modulation.

Nat Microbiol. 2025 Feb;10(2): 281-300

Design and regulation of engineered bacteria for environmental release.

Yonatan Chemla, Connor J Sweeney, Christopher A Wozniak, Christopher A Voigt.

Emerging products of biotechnology involve the release of living genetically modified microbes (GMMs) into the environment. However, regulatory challenges limit their use. So far, GMMs have mainly been tested in agriculture and environmental cleanup, with few approved for commercial purposes. Current government regulations do not sufficiently address modern genetic engineering and limit the potential of new applications, including living therapeutics, engineered living materials, self-healing infrastructure, anticorrosion coatings and consumer products. Here, based on 47 global studies on soil-released GMMs and laboratory microcosm experiments, we discuss the environmental behaviour of released bacteria and offer engineering strategies to help improve performance, control persistence and reduce risk. Furthermore, advanced technologies that improve GMM function and control, but lead to increases in regulatory scrutiny, are reviewed. Finally, we propose a new regulatory framework informed by recent data to maximize the benefits of GMMs and address risks.

DOI: https://doi.org/10.1038/s41564-024-01918-0
Nat Commun. 2025 Feb 08. 16(1): 1464

Synthetic transmembrane DNA receptors enable engineered sensing and actuation.

Ze-Rui Zhou, Man-Sha Wu, Zhenglin Yang, Yuting Wu, Weijie Guo, Da-Wei Li, Ruo-Can Qian, Yi Lu.

In living organisms, cells synergistically couple cascade reaction pathways to achieve inter- and intracellular signal transduction by transmembrane protein receptors. The construction and assembly of synthetic receptor analogs that can mimic such biological processes is a central goal of synthetic biochemistry and bionanotechnology to endow receptors with user-defined signal transduction effects. However, designing artificial transmembrane receptors with the desired input, output, and performance parameters are challenging. Here we show that the dimerization of synthetic transmembrane DNA receptors executes a systematically engineered sensing and actuation cascade in response to external molecular signals. The synthetic DNA receptors are composed of three parts, including an extracellular signal reception part, a lipophilic transmembrane anchoring part, and an intracellular signal output part. Upon the input of external signals, the DNA receptors can form dimers on the cell surface triggered by configuration changes, leading to a series of downstream cascade events including communication between donor and recipient cells, gene transcription regulation, protein level control, and cell apoptosis. We believe this work establishes a flexible cell surface engineering strategy that is broadly applicable to implement sophisticated biological functions.

DOI: https://doi.org/10.1038/s41467-025-56758-1
Curr Opin Biotechnol. 2025 Feb 06. pii: S0958-1669(25)00013-8. [Epub ahead of print]92 103269

Engineered microbial living matter for diagnostics, prevention, and therapy.

Ali Khazem, Rosanne Schmachtenberg, Anke Weiand, Shrikrishnan Sankaran, Wilfried Weber.

Living therapeutic and diagnostic materials based on engineered microorganisms are emerging as a novel approach with the perspective of providing patient-tailored, sustainable, and cost-effective healthcare solutions. In this review, we focus on recent advances in using genetically or chemically engineered microorganisms as living diagnostics, therapeutics, and as a means of prevention for various diseases. We also highlight the applications of living therapeutics for acute and chronic diseases, and the role of micro/macro-encapsulation of the engineered microorganisms. We further showcase the current success of engineered living therapeutics in clinical trials and discuss challenges and future trends in the field.

DOI: https://doi.org/10.1016/j.copbio.2025.103269
Biomater Sci. 2025 Feb 03.

A thiol-ene click-based strategy to customize injectable polymer-nanoparticle hydrogel properties for therapeutic delivery.

Sophia J Bailey, Noah Eckman, Elisa S Brunel, Carolyn K Jons, Samya Sen, Eric A Appel.

Polymer-nanoparticle (PNP) hydrogels are a promising injectable biomaterial platform that has been used for a wide range of biomedical applications including adhesion prevention, adoptive cell delivery, and controlled drug release. By tuning the chemical, mechanical, and erosion properties of injected hydrogel depots, additional control over cell compatibility and pharmaceutical release kinetics may be realized. Here, we employ thiol-ene click chemistry to prepare a library of modified hydroxypropylmethylcellulose (HPMC) derivatives for subsequent use in PNP hydrogel applications. When combined with poly(ethylene glycol)-b-poly(lactic acid) nanoparticles, we demonstrate that systematically altering the hydrophobic, steric, or pi stacking character of HPMC modifications can readily tailor the mechanical properties of PNP hydrogels. Additionally, we highlight the compatibility of the synthetic platform for the incorporation of cysteine-bearing peptides to access PNP hydrogels with improved bioactivity. Finally, through leveraging the tunable physical properties afforded by this method, we show hydrogel retention time in vivo can be dramatically altered without sacrificing mesh size or cargo diffusion rates. This work offers a route to optimize PNP hydrogels for a variety of translational applications and holds promise in the highly tunable delivery of pharmaceuticals and adoptive cells.

DOI: https://doi.org/10.1039/d4bm01315h
J Am Chem Soc. 2025 Feb 05.

Covalent Dynamic DNA Networks to Translate Multiple Inputs into Programmable Outputs.

Simone Brannetti, Serena Gentile, Erica Del Grosso, Sijbren Otto, Francesco Ricci.

Inspired by naturally occurring protein dimerization networks, in which a set of proteins interact with each other to achieve highly complex input-output behaviors, we demonstrate here a fully synthetic DNA-based dimerization network that enables highly programmable input-output computations. Our DNA-based dimerization network consists of DNA oligonucleotide monomers modified with reactive moieties that can covalently bond with each other to form dimer outputs in an all-to-all or many-to-many fashion. By designing DNA-based input strands that can specifically sequester DNA monomers, we can control the size of the reaction network and thus fine-tune the yield of each DNA dimer output in a predictable manner. Thanks to the programmability and specificity of DNA-DNA interactions, we show that this approach can be used to control the yield of different dimer outputs using different inputs. The approach is also versatile and we demonstrate dimerization networks based on two distinct covalent reactions: thiol-disulfide and strain-promoted azide-alkyne cycloaddition (SPAAC) reactions. Finally, we show here that the DNA-based dimerization network can be used to control the yield of a functional dimer output, ultimately controlling the assembly and disassembly of DNA nanostructures. The covalent dynamic DNA networks shown here provide a way to convert multiple inputs into programmable outputs that can control a broader range of functions, including ones that mimic those of living cells.

DOI: https://doi.org/10.1021/jacs.4c13854
Mater Horiz. 2025 Feb 03.

From 3D to 4D printing of lignin towards green materials and sustainable manufacturing.

Tingting Wu, Sigit Sugiarto, Ruochen Yang, Thenapakiam Sathasivam, Udyani Aloka Weerasinghe, Pei Lin Chee, Odelia Yap, Gustav Nyström, Dan Kai.

Lignin is the second most abundant renewable and sustainable biomass resource. Developing advanced manufacturing to process lignin/lignocellulose into functional materials could reduce the consumption of petroleum-based materials. 3D printing provides a promising strategy to realize complex and customized geometries of lignin materials. The heterogeneity and complexity of lignin hinder its processing via additive manufacturing, but the recent advancement in lignin modification and polymerization provides new opportunities. Here, we summarize the recent state-of-the-art 3D printing of lignin materials, including the selection and formulation of lignin materials based on different printing techniques, the chemical modification of lignin for enhanced printability, and the related application fields. Additionally, we highlight the significant role of the 3D printing of lignocellulose biomass materials, such as wood powder and agricultural wastes. It was concluded that the most challenging part is to enhance the printability of lignin materials through modification and pretreatment of lignin while keeping the whole process green and sustainable. Beyond 3D printing, we further discuss the development of smart lignin materials and their potential for 4D printing. Ultimately, we discuss the current challenges and potential opportunities for the additive manufacturing of lignin materials. We believe this review can raise awareness among researchers about the potential of lignin materials as whole materials for constructing blocks and can promote the development of 3D/4D printing of lignin towards sustainability.

DOI: https://doi.org/10.1039/d4mh01680g
ACS Appl Mater Interfaces. 2025 Feb 06.

Matrix Stiffness Regulates GBM Migration and Chemoradiotherapy Responses via Chromatin Condensation in 3D Viscoelastic Matrices.

Sauradeep Sinha, Mark Fleck, Manish Ayushman, Xinming Tong, Georgios Mikos, Sarah Jones, Luis Soto, Fan Yang.

  Glioblastoma multiforme (GBM) progression is associated with changes in matrix stiffness, and different regions of the tumor niche exhibit distinct stiffnesses. Using elastic hydrogels, previous work has demonstrated that matrix stiffness modulates GBM behavior and drug responses. However, brain tissue is viscoelastic, and how stiffness impacts the GBM invasive phenotype and response to therapy within a viscoelastic niche remains largely unclear. Here, we report a three-dimensional (3D) viscoelastic GBM hydrogel system that models the stiffness heterogeneity present within the tumor niche. We find that GBM cells exhibit enhanced migratory ability, proliferation, and resistance to radiation in soft matrices, mimicking the tumor core and perifocal margins. Conversely, GBM cells remain confined and demonstrate increased resistance to chemotherapy in stiff matrices mimicking edematous tumor regions. We identify that stiffness-induced changes in the GBM phenotype are regulated by nuclear mechanosensing and chromatin condensation. Pharmacologically decondensing the chromatin significantly impedes GBM migration and overcomes stiffness-induced chemoresistance and radioresistance. Our findings highlight that stiffness regulates aggressive GBM behavior in viscoelastic matrices through mechanotransduction processes. Finally, we reveal the critical role of chromatin condensation in mediating GBM migration and therapy resistance, offering a potential new therapeutic target to improve GBM treatment outcomes.

Keywords:  chromatin condensation; glioblastoma multiforme; mechanotransduction; stiffness; viscoelastic hydrogels

DOI:  https://doi.org/10.1021/acsami.4c16993
Macromol Rapid Commun. 2025 Feb 07. e2401156

Spatially Controlled Self-Assembly of Supramolecular Hydrogels Enabled by Light-Triggered Catalysis.

Jiahao Zhang, Kaiyu Jin, Yichen Xiao, Yifei Feng, Da Lu, Mai Chen, Mengran Sun, Dengyu Wang, Cheng Jin, Zhiling Li, Yiming Wang.

  Spatial control over supramolecular self-assembly prevails in living system, yet remains difficult to replicate in synthetic scenarios. Here, on the basis of a hydrazone formation-mediated supramolecular hydrogelation system, access to patterning of supramolecular hydrogels is demonstrated via a light-triggered catalysis strategy. A photoacid generator that can produce protons in aqueous solutions upon irradiation is employed. The generated protons lead to a drop in pH of around three units (initial pH 7.0), effectively accelerating the formation and self-assembly of the hydrazone gelators. Because of the light-triggered catalysis, the hydrogelation samples in the presence of photoacid generator show lower critical gelation concentration, higher stiffness, and denser networks. Importantly, by performing selective irradiation using differently shaped masks, various spatially resolved supramolecular hydrogels following the shapes of the masks are fabricated. The concept of using light-triggered catalysis to realize spatial control over supramolecular self-assembly provides an alternative approach toward bottom-up fabrication of structured soft materials for various applications such as tissue engineering, single cell manipulation, and biosensing.

Keywords:  directed self‐assembly; hydrogels; low‐molecular‐weight gelators; patterning; supramolecular chemistry

DOI:  https://doi.org/10.1002/marc.202401156
mBio. 2025 Feb 06. e0200823

A new flavor of synthetic yeast communities sees the light.

Vicente Rojas, Daniela Rivera, Carlos Ruiz, Luis F Larrondo.

  No organism is an island: organisms of varying taxonomic complexity, including genetic variants of a single species, can coexist in particular niches, cooperating for survival while simultaneously competing for environmental resources. In recent years, synthetic biology strategies have witnessed a surge of efforts focused on creating artificial microbial communities to tackle pressing questions about the complexity of natural systems and the interactions that underpin them. These engineered ecosystems depend on the number and nature of their members, allowing complex cell communication designs to recreate and create diverse interactions of interest. Due to its experimental simplicity, the budding yeast Saccharomyces cerevisiae has been harnessed to establish a mixture of varied cell populations with the potential to explore synthetic ecology, metabolic bioprocessing, biosensing, and pattern formation. Indeed, engineered yeast communities enable advanced molecule detection dynamics and logic operations. Here, we present a concise overview of the state-of-the-art, highlighting examples that exploit optogenetics to manipulate, through light stimulation, key yeast phenotypes at the community level, with unprecedented spatial and temporal regulation. Hence, we envision a bright future where the application of optogenetic approaches in synthetic communities (optoecology) illuminates the intricate dynamics of complex ecosystems and drives innovations in metabolic engineering strategies.

Keywords:  Saccharomyces cerevisiae; optoecology; optogenetics; synthetic biology; synthetic ecology; yeast

DOI:  https://doi.org/10.1128/mbio.02008-23
Nature. 2025 Feb 05.

H-bonded organic frameworks as ultrasound-programmable delivery platform.

Wenliang Wang, Yanshu Shi, Wenrui Chai, Kai Wing Kevin Tang, Ilya Pyatnitskiy, Yi Xie, Xiangping Liu, Weilong He, Jinmo Jeong, Ju-Chun Hsieh, Anakaren Romero Lozano, Brinkley Artman, Xi Shi, Nicole Hoefer, Binita Shrestha, Noah B Stern, Wei Zhou, David W McComb, Tyrone Porter, Graeme Henkelman, Banglin Chen, Huiliang Wang.

The precise control of mechanochemical activation within deep tissues using non-invasive ultrasound holds profound implications for advancing our understanding of fundamental biomedical sciences and revolutionizing disease treatments1-4. However, a theory-guided mechanoresponsive materials system with well-defined ultrasound activation has yet to be explored5,6. Here we present the concept of using porous hydrogen-bonded organic frameworks (HOFs) as toolkits for focused ultrasound (FUS) programmably triggered drug activation to control specific cellular events in the deep brain, through on-demand scission of the supramolecular interactions. A theoretical model is developed to potentially visualize the mechanochemical scission and ultrasound mechanics, providing valuable guidelines for the rational design of mechanoresponsive materials to achieve programmable control. To demonstrate the practicality of this approach, we encapsulate the designer drug clozapine N-oxide (CNO) into the optimal HOF nanocrystals for FUS-gated release to activate engineered G-protein-coupled receptors in the ventral tegmental area (VTA) of mice and rats and hence achieve targeted neural circuit modulation even at depth 9 mm with a latency of seconds. This work demonstrates the capability of ultrasound to precisely control molecular interactions and develops ultrasound-programmable HOFs to non-invasively and spatiotemporally control cellular events, thereby facilitating the establishment of precise molecular therapeutic possibilities.

DOI: https://doi.org/10.1038/s41586-024-08401-0
Biomacromolecules. 2025 Feb 07.

3D-Printed Protein-Based Bioplastics with Tunable Mechanical Properties Using Glycerol or Hyperbranched Poly(glycerol)s as Plasticizers.

S Cem Millik, Naroa Sadaba, Shayna L Hilburg, Eva Sanchez-Rexach, Meijing Zhang, Siwei Yu, Alexander F Vass, Lilo D Pozzo, Alshakim Nelson.

Protein-based materials can be engineered to derive utility from the structures and functions of the incorporated proteins. Modern methods of protein engineering bring promise of unprecedented control over molecular and network design, which will enable new and improved functionalities in materials that incorporate proteins as functional building blocks. For these advantages to be fully realized, there is a need for robust methods for producing protein-based networks, as well as methods for tuning their mechanical properties. Light-based 3D-printing techniques afford high-resolution fabrication capability with unparalleled design freedom in an inexpensive and decentralized capacity. This work features 3D-printed serum albumin-based bioplastics with mechanical properties modulated through the incorporation of glycerol or hyperbranched poly(glycerol)s (HPGs) as plasticizers. These materials capitalize upon important features of serum albumin, including its low intrinsic viscosity, high aqueous solubility, and relatively low cost. The incorporation of glycerol or HPGs of different sizes resulted in softer and more ductile bioplastics than those obtained natively without additives. These bioplastics showed shape-memory behavior and could be used to fabricate functional objects. These materials are accessible, possess minimal chemical hazards, and can be used for fabricating rigid and strong as well as soft and ductile parts using inexpensive commercial 3D printers.

DOI: https://doi.org/10.1021/acs.biomac.4c01497
Nature. 2025 Feb 05.

Engineering a genomically recoded organism with one stop codon.

Michael W Grome, Michael T A Nguyen, Daniel W Moonan, Kyle Mohler, Kebron Gurara, Shenqi Wang, Colin Hemez, Benjamin J Stenton, Yunteng Cao, Felix Radford, Maya Kornaj, Jaymin Patel, Maisha Prome, Svetlana Rogulina, David Sozanski, Jesse Tordoff, Jesse Rinehart, Farren J Isaacs.

The genetic code is conserved across all domains of life, yet exceptions have revealed variations in codon assignments and associated translation factors1-3. Inspired by this natural malleability, synthetic approaches have demonstrated whole-genome replacement of synonymous codons to construct genomically recoded organisms (GROs)4,5 with alternative genetic codes. However, no efforts have fully leveraged translation factor plasticity and codon degeneracy to compress translation function to a single codon and assess the possibility of a non-degenerate code. Here we describe construction and characterization of Ochre, a GRO that fully compresses a translational function into a single codon. We replaced 1,195 TGA stop codons with the synonymous TAA in ∆TAG Escherichia coli C321.∆A4. We then engineered release factor 2 (RF2) and tRNATrp to mitigate native UGA recognition, translationally isolating four codons for non-degenerate functions. Ochre thus utilizes UAA as the sole stop codon, with UGG encoding tryptophan and UAG and UGA reassigned for multi-site incorporation of two distinct non-standard amino acids into single proteins with more than 99% accuracy. Ochre fully compresses degenerate stop codons into a single codon and represents an important step toward a 64-codon non-degenerate code that will enable precise production of multi-functional synthetic proteins with unnatural encoded chemistries and broad utility in biotechnology and biotherapeutics.

DOI: https://doi.org/10.1038/s41586-024-08501-x
Mater Today Bio. 2025 Apr;31 101472

Rapid isolation and recovery of Salmonella using hollow glass microspheres coated with multilayered nanofilms.

Rutwik Joshi, Hesaneh Ahmadi, Md Nayeem Hasan Kashem, Fariha Afnan, Siva Parameswaran, Chau-Chyun Chen, Gizem Levent, Wei Li.

Timely isolation, recovery, and identification of Salmonella from food samples is essential for prevention and control of foodborne Salmonella outbreaks. Traditional culture-based Salmonella isolation and serotyping techniques are time consuming and labor intensive. Despite the progress of innovative microfluidic or immunomagnetic isolation techniques, sophisticated lab equipment and microfabrication are often needed. Here, we present a novel, rapid yet simple method for isolation and recovery of Salmonella from mixed bacterial populations in food matrices and blood. This method utilizes self-floating hollow glass microspheres (HGMS) coated with biodegradable layer-by-layer (LbL) films and Salmonella specific antibodies. The isolation and recovery process can be completed in less than 2 h, without any sophisticated laboratory equipment or external force. In this study, we demonstrate that Salmonella can be captured due to antigen-antibody interactions on the surface of HGMS, allowing them to float to the top. The HGMS can then be washed and subjected to enzymatic degradation of the LbL film to recover the captured bacteria. The recovered Salmonella can subsequently be grown on selective agar plates for further analysis. Recovery efficiency of up to 22 % and detection limit of 100 CFU/mL were achieved. This method is expected to provide a viable alternative to traditional isolation techniques, especially in resource limited areas.

DOI: https://doi.org/10.1016/j.mtbio.2025.101472
Curr Opin Biotechnol. 2025 Feb 04. pii: S0958-1669(25)00009-6. [Epub ahead of print]92 103265

Harnessing photosynthesis for materials, devices, and environmental technologies.

Fan Jiang, Friedrich H Kleiner, Marie-Eve Aubin-Tam.

Photosynthetic organisms convert solar light into chemical energy through the process of photosynthesis. The employment of photosynthetic organisms in novel materials and devices provides them with a solar-powered and sustainable functionality. In general, photosynthesis utilizes light, water, and CO2 to generate various organic compounds while releasing secondary valuable products such as O2, extracellular electrons, carbohydrates, or H2. The light-dependent inputs and outputs are harnessed for environmental purification, biomedical applications, and production of biofuel, electricity, nanomaterials, or bioplastics. In this review, we summarize photosynthesis-assisted materials and engineering applications based on the products and substrates of photosynthetic processes, and we highlight key challenges that remain to be addressed.

DOI: https://doi.org/10.1016/j.copbio.2025.103265
J Control Release. 2025 Jan 31. pii: S0168-3659(25)00095-1. [Epub ahead of print]379 944-950

Immobilization of BMP-2 in porous hydrogels to spatially regulate osteogenesis.

Junzhe Lou, Charlotte Meyer, Anqi Chen, David A Weitz, David J Mooney.

  Sustained release of bone morphogenetic protein 2 (BMP-2) is used to enhance bone regeneration, but immobilizing BMP-2 in three-dimensional scaffolds could enable spatial regulation of stem cell differentiation and bone formation. Here, we fabricate porous granular hydrogels presenting BMP-2 on the surface to regulate stem cell growth and differentiation. Immobilization of BMP-2 and cell-adhesive ligands is achieved by surface-specific functionalization of microgels, which are jammed to form microporous hydrogels. Varying surface ligand density regulated spreading, proliferation and differentiation of cells. In addition, modulating the distribution of cell-adhesive ligands and BMP-2 allowed spatial control over cell adhesion and osteogenic differentiation.

Keywords:  BMP-2 immobilization; Granular hydrogel; Osteogenic differentiation; Spatial control

DOI:  https://doi.org/10.1016/j.jconrel.2025.01.084
Biomaterials. 2025 Jan 28. pii: S0142-9612(25)00075-4. [Epub ahead of print]318 123156

Bioprinting spatially guided functional 3D neural circuits with agarose-xanthan gum copolymer hydrogels.

Cristina Antich, Srikanya Kundu, Shayne Frebert, Ty Voss, Min Jae Song, Marc Ferrer.

  Engineered three-dimensional (3D) tissue models are being used as predictive human in vitro assays for drug discovery and development. Tissue engineering technologies such as bioprinting are now available which use mixtures of polymeric hydrogels and cells for the construction of biomimetic engineered 3D tissue models. Many of the polymeric hydrogels used for bioprinting require post-printing processing steps which might hinder their application directly in multi-well plate platforms, thus limiting their utility in a drug screening setting. Here we describe an agarose and xanthan gum copolymer hydrogel (AG-XG) that has optimal rheological properties for high shape fidelity with extrusion-based printing, has long term stability in cell culture conditions, and is "ready-to-use" after printing, not requiring post-printing processing treatments, making it ideal for applications in multi-well plate format. This AG-XG hydrogel is non-degradable and has non-cell permissive features which makes it ideal to create customized spatially guided cellular patterns to enhance relevant tissue geometry and function. As a proof-of-concept, we show that a bioprinted AG-XG hydrogel casting mold significantly enhances functional connectivity of an engineered 3D neural circuit model made using human iPSC-derived GABAergic and dopaminergic neurons and astrocytes. The bioprinted AG-XG mold promotes the formation of strong functional synaptic connections between two spatially separated neuronal regions, as measured with calcium and optogenetic-based fluorescent biosensors with a customized fiber photometry device. The high shape fidelity of the AG-XG hydrogels described here enables the biofabrication of precisely positioned and spatially designed cellular models, in muti well-based platforms used for drug screening. The process of printing these AG-XG hydrogels uses commercially available extrusion-based bioprinters and can therefore be easily implemented in translational laboratories doing tissue modeling and drug screening without the need of additional specialized bioengineering equipment.

Keywords:  3D cellular patterning; Agarose; Copolymeric hydrogel; Drug screening; Extrusion-based bioprinting; Multi-well plate format; Neural circuits; Tissue engineering; Tissue models; Xanthan gum

DOI:  https://doi.org/10.1016/j.biomaterials.2025.123156
Acc Chem Res. 2025 Feb 05.

Biological Polymers: Evolution, Function, and Significance.

Kavita Matange, Eliav Marland, Moran Frenkel-Pinter, Loren Dean Williams.

ConspectusA holistic description of biopolymers and their evolutionary origins will contribute to our understanding of biochemistry, biology, the origins of life, and signatures of life outside our planet. While biopolymer sequences evolve through known Darwinian processes, the origins of the backbones of polypeptides, polynucleotides, and polyglycans are less certain. We frame this topic through two questions: (i) Do the characteristics of biopolymer backbones indicate evolutionary origins? (ii) Are there reasonable mechanistic models of such pre-Darwinian evolutionary processes? To address these questions, we have established criteria to distinguish chemical species produced by evolutionary mechanisms from those formed by nonevolutionary physical, chemical, or geological processes. We compile and evaluate properties shared by all biopolymer backbones rather than isolating a single type. Polypeptide, polynucleotide, and polyglycan backbones are kinetically trapped and thermodynamically unstable in aqueous media. Each biopolymer forms a variety of elaborate assemblies with diverse functions, a phenomenon we call polyfunction. Each backbone changes structure and function upon subtle chemical changes such as the reduction of ribose or a change in the linkage site or stereochemistry of polymerized glucose, a phenomenon we call function-switching. Biopolymers display homo- and heterocomplementarity, enabling atomic-level control of structure and function. Biopolymer backbones access recalcitrant states, where assembly modulates kinetics and thermodynamics of hydrolysis. Biopolymers are emergent; the properties of biological building blocks change significantly upon polymerization. In cells, biopolymers compose mutualistic networks; a cell is an Amazon Jungle of molecules. We conclude that biopolymer backbones exhibit hallmarks of evolution. Neither chemical, physical, nor geological processes can produce molecules consistent with observations. We are faced with the paradox that Darwinian evolution relies on evolved backbones but cannot alter biopolymer backbones. This Darwinian constraint is underlined by the observation that across the tree of life, ribosomes are everywhere and always have been composed of RNA and protein. Our data suggest that chemical species on the Hadean Earth underwent non-Darwinian coevolution driven in part by hydrolytic stress, ultimately leading to biopolymer backbones. We argue that highly evolved biopolymer backbones facilitated a seamless transition from chemical to Darwinian evolution. This model challenges convention, where backbones are products of direct prebiotic synthesis. In conventional models, biopolymer backbones retain vestiges of prebiotic chemistry. Our findings, however, align with models where chemical species underwent iterative and recursive sculpting, selection, and exaptation. This model supports Orgel's "gloomy" prediction that modern biochemistry has discarded vestiges of prebiotic chemistry. But there is hope. We believe an understanding of biopolymer origins will progress during the challenging and exciting integration of chemical sciences and evolutionary theory. These efforts can provide new perspectives on pre-Darwinian mechanisms and can deepen our understanding of evolution and of chemical sciences. Our working definition of chemical evolution is continuous chemical change with exploration of new chemical spaces and avoidance of equilibrium. In alignment with our model, we observe chemical evolution in complex mixtures undergoing wet-dry cycling, which does appear to undergo continuous chemical change and exploration of new chemical spaces while avoiding equilibrium.

DOI: https://doi.org/10.1021/acs.accounts.4c00546
Mater Today Bio. 2025 Apr;31 101452

Surface functionalization of microscaffolds produced by high-resolution 3D printing: A new layer of freedom.

Oliver Kopinski-Grünwald, Stephan Schandl, Jegor Gusev, Ourania Evangelia Chamalaki, Aleksandr Ovsianikov.

  Scaffolded-spheroids represent novel building blocks for bottom-up tissue assembly, allowing to produce constructs with high initial cell density. Previously, we demonstrated the successful differentiation of such building blocks, produced from immortalized human adipose-derived stem cells, towards different phenotypes, and the possibility of creating macro-sized tissue-like constructs in vitro. The culture of cells in vitro depends on the supply of various nutrients and biomolecules, such as growth factors, usually supplemented in the culture medium. Another means for growth factor delivery (in vitro and in vivo) is the release from the scaffold to alter the biological response of surrounding cells (e.g. by release of VEGF).1 As a proof of concept for this approach, we sought to biofunctionalize the surface of the microscaffolds with heparin as a "universal linker" that would allow binding a variety of growth factors/biomolecules. An aminolysis step in an organic solvent made it possible to generate a hydrophilic and charged surface. The backbone of the amine, as well as reaction conditions, led to an adjustable surface modification. The amount of heparin on the surface was increased with an ethylene glycol-based diamine backbone and varied between 8 and 40 ng per microscaffold. Choosing a suitable linker allows easy adjustment of the loading of VEGF and other heparin-binding proteins. Initial results indicated that up to 5 ng VEGF could be loaded per microscaffold, generating a steady VEGF release for 16 days. We report an easy-to-perform, scalable surface modification approach of polyester-based resin that leads to adjustable surface concentrations of heparin. The successful surface aminolysis opens the route to various modifications and broadens the spectrum of biomolecules which can be delivered.

Keywords:  Growth factors; High-resolution 3D printing; Microscaffolds; Scaffolded spheroids; Surface modification; Tissue engineering; VEGF

DOI:  https://doi.org/10.1016/j.mtbio.2025.101452
Small. 2025 Feb 05. e2406388

Proteins for Hyperelastic Materials.

Rui Su, Chao Ma, Bing Han, Hongjie Zhang, Kai Liu.

  Meticulous engineering and the yielded hyperelastic performance of structural proteins represent a new frontier in developing next-generation functional biomaterials. These materials exhibit outstanding and programmable mechanical properties, including elasticity, resilience, toughness, and active biological characteristics, such as degradability and tissue repairability, compared with their chemically synthetic counterparts. However, there are several critical issues regarding the preparation approaches of hyperelastic protein-based materials: limited natural sequence modules, non-hierarchical assembly, and imbalance between compressive and tensile elasticity, leading to unmet demands. Therefore, it is pivotal to develop an alternative strategy for biofabricating hyperelastic materials. Herein, the molecular design, engineering, and property regulation of hyperelastic structural proteins are overviewed. First, methodologies for deeper exploration of mechanical modules, including machine learning-aided de novo design, random mutations of natural sequences, and multiblock fusion techniques, are actively introduced. These methodologies facilitate the generation of elastomeric protein modules and demonstrate enhanced structural and functional versatility. Subsequently, assembly tactics of hyperelastic proteins (i.e., physical modulation, genetic adaptations, and chemical modifications) are reviewed, yielding hierarchically ordered structures. Finally, advances in biophysical techniques for more nuanced characterization of protein ensembles are discussed, unveiling the tuning mechanisms of protein elasticity across scales. Future developments in structural hyperelastic protein-based biomaterials are also envisioned.

Keywords:  engineering; hyperelastic material; molecular design; performance regulation; structural protein

DOI:  https://doi.org/10.1002/smll.202406388
ACS Synth Biol. 2025 Feb 04.

Synthetic Dual-Input Hybrid Riboswitches─Optimized Genetic Regulators in Yeast.

Daniel Kelvin, Janette Arias Rodriguez, Ann-Christin Groher, Kiara Petras, Beatrix Suess.

  Synthetic riboswitches, genetic regulatory elements composed entirely of RNA, have been engineered to control a variety of mechanisms at the level of both transcription and translation in all domains of life. The efficiency of riboswitch regulation can be increased by inserting two of them into an mRNA sequence in close proximity, resulting in a tandem riboswitch. The tandem state results in improved regulation beyond that of a single riboswitch by allowing both binding pockets to contribute to a higher dynamic range. The focus of this study was to create a novel tandem riboswitch design by integrating the binding pockets of two different riboswitches into one continuous structure, thereby creating a dual-input hybrid riboswitch. These hybrids remain compact in size with a shorter sequence length compared to a tandem riboswitch, while taking advantage of the binding pockets and scaffold sequences provided by both parental riboswitches. Through rational design, hybrid constructs derived from the combination of tetracycline-, tobramycin-, neomycin-, and paromomycin-binding riboswitches were engineered that significantly increase the dynamic range (e.g., from 14- to 36-fold for tobramycin) while increasing their expression levels in the absence of ligand (e.g., 28% to 68% expression for tetracycline). This study expands the toolbox of synthetic riboswitches and establishes general design guidelines applicable to similar riboswitches. Additionally, the dual-input state makes hybrid riboswitches an interesting target for the design of genetic regulators following Boolean logic.

Keywords:  aptamer; dual input; hybrid riboswitch; riboswitch; synthetic biology; tandem constructs

DOI:  https://doi.org/10.1021/acssynbio.4c00660
Adv Mater. 2025 Feb 05. e2416260

Engineered Living Systems Based on Gelatin: Design, Manufacturing, and Applications.

Zhenwu Wang, Zeng Lin, Xuan Mei, Ling Cai, Ko-Chih Lin, Jimena Flores Rodríguez, Zixin Ye, Ximena Salazar Parraguez, Emilio Mireles Guajardo, Pedro Cortés García Luna, Jun Yi Joey Zhang, Yu Shrike Zhang.

  Engineered living systems (ELSs) represent purpose-driven assemblies of living components, encompassing cells, biomaterials, and active agents, intricately designed to fulfill diverse biomedical applications. Gelatin and its derivatives have been used extensively in ELSs owing to their mature translational pathways, favorable biological properties, and adjustable physicochemical characteristics. This review explores the intersection of gelatin and its derivatives with fabrication techniques, offering a comprehensive examination of their synergistic potential in creating ELSs for various applications in biomedicine. It offers a deep dive into gelatin, including its structures and production, sources, processing, and properties. Additionally, the review explores various fabrication techniques employing gelatin and its derivatives, including generic fabrication techniques, microfluidics, and various 3D printing methods. Furthermore, it discusses the applications of ELSs based on gelatin in regenerative engineering as well as in cell therapies, bioadhesives, biorobots, and biosensors. Future directions and challenges in gelatin fabrication are also examined, highlighting emerging trends and potential areas for improvements and innovations. In summary, this comprehensive review underscores the significance of gelatin-based ELSs in advancing biomedical engineering and lays the groundwork for guiding future research and developments within the field.

Keywords:  3D printing; biofabrication; engineered living systems; gelatin; hydrogels

DOI:  https://doi.org/10.1002/adma.202416260
ACS Synth Biol. 2025 Feb 04.

DRIMS: A Synthetic Biology Platform that Enables Deletion, Replacement, Insertion, Mutagenesis, and Synthesis of DNA.

Leidy D Caraballo G, Inci Cevher Zeytin, Purva Rathi, Che-Hsing Li, Ai-Ni Tsao, Yaery J Salvador L, Manish Ranjan, Brendan Magee Traynor, Andras A Heczey.

  DNA modification and synthesis are fundamental to genetic engineering, and systems that enable time- and cost-effective execution of these processes are crucial. Iteration of genetic construct variants takes significant time, cost and effort to develop new therapeutic strategies to treat diseases including cancer. Thus, decreasing cost and enhancing simplicity while accelerating the speed of advancement is critical. We have developed a PCR-based platform that allows for deletion, replacement, insertion, mutagenesis, and synthesis of DNA (DRIMS). These modifications rely on the recA-independent recombination pathway and are carried out in a single amplification step followed by DpnI digestion and transformation into competent cells. DNA synthesis is accomplished through sequential PCR amplification reactions without the need for a DNA template. Here, we provide proof-of-concept for the DRIMS platform by performing four deletions within DNA fragments of various sizes, sixty-four replacements of DNA binding sequences that incorporate repeat sequences, four replacements of chimeric antigen receptor components, fifty-one insertions of artificial microRNAs that form complex tertiary structures, five varieties of point mutations, and synthesis of eight DNA sequences including two with high GC content. Compared to other advanced cloning methods including Gibson and "in vivo assembly", we demonstrate the significant advantages of the DRIMS platform. In summary, DRIMS allows for efficient modification and synthesis of DNA in a simple, rapid and cost-effective manner to accelerate the synthetic biology field and development of therapeutics.

Keywords:  DNA cloning; DNA synthesis; PCR; chimeric antigen receptor (CAR); genetic engineering; mutagenesis

DOI:  https://doi.org/10.1021/acssynbio.4c00649
bioRxiv. 2025 Jan 24. pii: 2025.01.23.634604. [Epub ahead of print]

Engineering microalgal cell wall-anchored proteins using GP1 PPSPX motifs and releasing with intein-mediated fusion.

Kalisa Kang, Évellin do Espirito Santo, Crisandra Jade Diaz, Stephen Mayfield, João Vitor Dutra Molino.

Harnessing and controlling the localization of recombinant proteins is critical for advancing applications in synthetic biology, industrial biotechnology, and drug delivery. This study explores protein anchoring and controlled release in Chlamydomonas reinhardtii , providing innovative tools for these fields. Using truncated variants of the GP1 glycoprotein fused to the plastic-degrading enzyme PHL7, we identified the PPSPX motif as essential for anchoring proteins to the cell wall. Constructs with increased PPSPX content exhibited reduced secretion but improved anchoring, pinpointing the potential anchor-signal sites of GP1 and highlighting the distinct roles of these motifs in protein localization. Building on the anchoring capabilities established with these glycomodules, we also demonstrated a controlled release system using a pH-sensitive intein derived from RecA from Mycobacterium tuberculosis . This intein efficiently cleaved and released PHL7 and mCherry that was fused to GP1 under acidic conditions, enabling precise temporal and environmental control. At pH 5.5, fluorescence kinetics demonstrated significant mCherry release from the pJPW4mCherry construct within 4 hours. In contrast, release was minimal under pH 8.0 conditions and negligible for the pJPW2mCherry (W2) control, irrespective of the pH. Additionally, bands on the Western blot at the expected size of mCherry also showed its efficient release from the mCherry::intein::GP1 fusion protein at pH 5.5. Conversely, at pH 8.0, no bands were detected. This anchor-release approach offers significant potential for drug delivery, biocatalysis, and environmental monitoring applications. By integrating glycomodules and pH-sensitive inteins, this study establishes a versatile framework for optimizing protein localization and release in C. reinhardtii , with broad implications for proteomics, biofilm engineering, and scalable therapeutic delivery systems.

DOI: https://doi.org/10.1101/2025.01.23.634604
Proc Natl Acad Sci U S A. 2025 Feb 11. 122(6): e2417058122

Spatial propagation of temperate phages within and among biofilms.

James B Winans, Lanying Zeng, Carey D Nadell.

  Bacteria form groups composed of cells and a secreted polymeric matrix that controls their spatial organization. These groups-termed biofilms-can act as refuges from environmental disturbances and from biotic threats, including phages. Despite the ubiquity of temperate phages and bacterial biofilms, live propagation of temperate phages within biofilms has not been characterized on cellular spatial scales. Here, we leverage several approaches to track temperate phages and distinguish between lytic and lysogenic host infections. We determine that lysogeny within Escherichia coli biofilms initially occurs within a predictable region of cell group packing architecture on the biofilm periphery. Because lysogens are generally found on the periphery of large cell groups, where lytic viral infections also reduce local biofilm structural integrity, lysogens are predisposed to disperse into the passing liquid and are overrepresented in downstream biofilms formed from the dispersal pool of the original biofilm-phage system. Comparing our results with those for virulent phages reveals that temperate phages have unique advantages in propagating over long spatial and time scales within and among bacterial biofilms.

Keywords:  biofilm; dispersal; matrix; phage; spatial ecology

DOI:  https://doi.org/10.1073/pnas.2417058122
Nat Chem Biol. 2025 Feb 05.

Programming biological communication between distinct membraneless compartments.

Bo-Tao Ji, He-Tong Pan, Zhi-Gang Qian, Xiao-Xia Xia.

Distinct membraneless organelles within cells collaborate closely to organize crucial functions. However, biosynthetic communicating membraneless organelles have yet to be created. Here we report a binary population of membraneless compartments capable of coexistence, biological communication and controllable feedback under cellular environmental conditions. The compartment consortia emerge from two orthogonally phase-separating proteins in a cell-free expression system. Their appearance can be programmed in time and order for on-demand delivery of molecules. In particular, the consortia can sense, process and deliver functional protein cargo in response to a protease message or a DNA message that encodes the protease. Such DNA-based molecular programs can be further harnessed by installing a feedback loop that controls the information flow at the messenger RNA level. These results contribute to understanding crosstalk among membraneless organelles and provide a design principle that can guide construction of functional compartment consortia.

DOI: https://doi.org/10.1038/s41589-025-01840-4
Biomater Sci. 2025 Feb 06.

Crosslink strength governs yielding behavior in dynamically crosslinked hydrogels.

Noah Eckman, Abigail K Grosskopf, Grace Jiang, Krutarth Kamani, Michelle S Huang, Brigitte Schmittlein, Sarah C Heilshorn, Simon Rogers, Eric A Appel.

Yielding of dynamically crosslinked hydrogels, or the transition between a solid-like and liquid-like state, allows facile injection and utility in translational biomedical applications including delivery of therapeutic cells. Unfortunately, the time-varying nature of the transition is not well understood, nor are there design rules for understanding the effects of yielding on encapsulated cells. Here, we unveil underlying molecular mechanisms governing the yielding transition of dynamically crosslinked gels currently being researched for use in cell therapy. We demonstrate through nonlinear rheological characterization that the network dynamics of the dynamic hydrogels dictate the speed and character of their yielding transition. Rheological testing of these materials reveals unexpected elastic strain stiffening during yielding, as well as characterization of the rapidity of the yielding transition. A slower yielding speed explains enhanced protection of directly injected cells from shear forces, highlighting the importance of mechanical characterization of all phases of yield-stress biomaterials.

DOI: https://doi.org/10.1039/d4bm01323a
Heliyon. 2025 Jan 30. 11(2): e41993

New trends of additive manufacturing using materials based-on natural fibers and minerals : A systematic review.

Joao Ribeiro, Manuel Rodríguez-Martín, Joaquín Barreiro, Ana Fernández-Abia, Roberto García-Martín, Joao Rocha, Susana Martínez-Pellitero.

  Polymeric materials based on natural fibers and minerals are currently being researched and their development is still in its infancy but is expected to increase in the coming years (being nowadays a hot topic). Their main advantage is that they make it possible to use waste and by-products of agricultural, forestry, and mineral origin to generate materials for Additive Manufacturing. Since their use reduces the need for other synthetic polymers derived from petroleum and other non-natural fibers that generate a high environmental impact, this type of material is a sustainable, environmentally friendly, biodegradable solution that can be integrated into the value chain of certain industries and, finally, favors the circular economy. This study presents a bibliometric analysis, meta-analysis, and systematic literature review focusing on plant-based fibers and minerals in biocomposites from a holistic perspective. To learn about the potential of these new materials at an industrial level and to learn about the benefits they can have for society, the strengths, weaknesses, opportunities, and threats have been evaluated. The results strongly suggest that these materials will undergo intensive development in the upcoming years, with a substantial increase in their integration across industries.

Keywords:  Additive manufacturing (AM); Composites; Manufacturing processes; Mineral additives; Natural fibers

DOI:  https://doi.org/10.1016/j.heliyon.2025.e41993
Sci Rep. 2025 Feb 04. 15(1): 4213

A polysaccharide-based hydrogel platform for tumor spheroid production and anticancer drug screening.

Elliot Lopez-Vince, Teresa Simon-Yarza, Claire Wilhelm.

  Extracellular matrix mimics are still needed to grow cancer cells in 3D environments and study their evolution in vitro while precisely controlling relevant features. Most models currently use collagen, which is biomimetic but degrades quickly, or artificial polymers, which can be chemically modified but remain stiff. Herein we introduced a soft, non-adhesive, and resistant hydrogel platform for tumor spheroid production using a polysaccharide-based formulation. To ensure micro-structuring of the hydrogel and enable spheroid formation, 3D printed molds consisting of a network of 200-µm-diameter micropillars were used to generate microstructured hydrogel constructs that fit into a multi-well plate. This platform was validated for drug testing using three cancer cell lines (A673, MCF7 and U87) and 2 anticancer drugs (doxorubicin and paclitaxel). Drug response was assessed through bright-field microscopy monitoring and viability measurements after 48 h of treatment. This study validates the use of pullulan-dextran hydrogels for spheroid formation, combined with in situ drug screening.

Keywords:  Biomaterials; Drug screening; Hydrogels; In vitro models; Spheroids; Tumor models

DOI:  https://doi.org/10.1038/s41598-025-87896-7
Appl Environ Microbiol. 2025 Feb 07. e0192624

Polysulfides promote protein disulfide bond formation in microorganisms growing under anaerobic conditions.

Yuping Xin, Qingda Wang, Jianming Yang, Xiaohua Wu, Yongzhen Xia, Luying Xun, Huaiwei Liu.

  Polysulfides commonly occur in anaerobic, microbial active environments, where they play key roles in sulfur cycling and redox transformations. Anaerobic survival of microorganisms requires the formation of protein disulfide bond (DSB). The relationship between polysulfides and anaerobic DSB formation has not been studied so far. Herein, we discovered that polysulfides can efficiently mediate protein DSB formation of microorganisms under anaerobic conditions. We used polysulfides to treat proteins, including roGFP2, Trx1, and DsbA, under anaerobic conditions and found that all three proteins formed intramolecular DSB in vitro. Under anaerobic conditions, Escherichia coli ΔdsbB displayed reduced growth and decreased intracellular protein DSB levels, but polysulfide treatment restored both growth and DSB content. Similarly, polysulfide treatment of E. coli ΔdsbA promoted periplasmic roGFP2 DSB formation and recovered growth under anaerobic conditions. Furthermore, treating Schizosaccharomyces pombe and Cupriavidus pinatubonensis JMP134 with polysulfides increased their intracellular protein DSB content. Collectively, these findings demonstrate that polysulfides can promote DSB formation independently of known enzymatic DSB-mediated systems and the presence of oxygen, thereby benefiting the survival of microorganisms in anaerobic habitats.IMPORTANCEHow polysulfides enhance the adaption of microorganisms to anaerobic environments remains unclear. Our study reveals that polysulfides efficiently facilitate protein DSB formation under anaerobic conditions. Polysulfides contain zero-valent sulfur atoms (S0), which can be transferred to the thiol group of cysteine residue. This S0 atom then accepts two electrons from two cysteine residues and is reduced to H2S, leaving the two cysteines linked by a disulfide bond. Anaerobic growth of microorganisms benefits from the formation of DSB. These findings pave the way for a deeper understanding of the intricate relationship between polysulfides and microorganisms in various environmental contexts.

Keywords:  DSB system; anaerobic growth; polysulfides; protein disulfide bond; s-glutathionylation

DOI:  https://doi.org/10.1128/aem.01926-24
Curr Opin Chem Biol. 2025 Feb 03. pii: S1367-5931(25)00001-8. [Epub ahead of print]85 102569

Optogenetic engineering for ion channel modulation.

Tianlu Wang, Tatsuki Nonomura, Tien-Hung Lan, Yubin Zhou.

  Optogenetics, which integrates photonics and genetic engineering to control protein activity and cellular processes, has transformed biomedical research. Its precise spatiotemporal control, minimal invasiveness, and tunable reversibility have spurred its widespread adoption in both basic and clinical research. Optogenetic techniques have been applied to partially restore vision in blind patients and are being actively explored as innovative treatments for neurological, psychiatric, cardiac, and immunological disorders. Microbial channelrhodopsins (ChRs) allow precise manipulation of neuronal and cardiac activities, while vertebrate rhodopsins offer unique opportunities for ion channel modulation through G-protein-coupled receptor (GPCR) pathways. Plant-derived photoswitchable domains can also be engineered into ion channels to confer photosensitivity. This review summarizes the latest progress in engineering genetically encoded light-sensitive ion channel actuators and modulators (GELICAMs) with diverse ion selectivity and spectral sensitivity. We further discuss the potential applications and challenges of these tools in advancing biomedical research and therapeutic interventions.

Keywords:  Channelrhodopsin; Immunotherapy; Ion channel; Neurodegeneration; Optogenetics; Protein engineering; Synthetic biology

DOI:  https://doi.org/10.1016/j.cbpa.2025.102569