2026-05-03T08:20:52Z https://api.figshare.com/v2/oai

oai:figshare.com:article/32147880 2026-05-02T00:00:00Z category_1 category_4 category_7 category_12 category_21 category_61 category_734 portal_63 item_type_6 month_year_05_2026

Covalent Anchoring of Thymol/Zein Nanoparticles on Wool Fibers via ε‑Polylysine Cross-linking for Durable Antibacterial Performance Yujie Guo (4760289) Jing Meng (586670) Jiangman Ma (23827539) Hongli Zhu (1419181) Qinfei Ke (1622527) Yi Zhao (14034) Biophysics Biochemistry Medicine Cell Biology Biotechnology Developmental Biology Biological Sciences not elsewhere classified xps analyses confirmed synergistic antibacterial activity poses potential risk lasting antibacterial activity forming amide bonds conventional physical adsorption based fiber materials bacterial contamination owing antisolvent coprecipitation method acyl transfer reactions natural polypeptide ε work presents wool fibers wool felt thermal stability textile industry significantly shortens service life proteinaceous composition pl ). markedly enhanced innovative enzyme human health highly susceptible functional protein effectively overcoming covalent anchoring continuous coating average size 129 nm 120 h Wool is one of the most widely used raw materials in the textile industry. However, it is highly susceptible to bacterial contamination owing to proteinaceous composition, which significantly shortens its service life and poses potential risk to human health. Herein, we prepare thymol-loaded zein nanoparticles via an antisolvent coprecipitation method, with the natural polypeptide ε-polylysine (ε-PLL) serving as an interfacial cross-linking bridge between the microcapsules and wool fibers. The nanoparticles exhibited uniform spherical morphology with an average size of 129 nm. FTIR and XPS analyses confirmed that ε-PLL was cross-linked with nanoparticles and wool fibers through acyl transfer reactions, forming amide bonds, which markedly enhanced the thermal stability of the system. Morphology and pore size analyses revealed that ε-PLL interfacially bridged nanoparticle modification produced a continuous coating on the surface of the wool felt (OW-Z@PL). Under the synergistic antibacterial activity of ε-polylysine and thymol, OW-Z@PL exhibited highly efficient and long-lasting antibacterial activity, maintaining high antibacterial performance even after 120 h and effectively overcoming the limitations of conventional physical adsorption. This work presents an innovative enzyme-catalyzed covalent modification strategy for the development of green, durable, and functional protein-based fiber materials. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acssuschemeng.6c01493.s001 https://figshare.com/articles/journal_contribution/Covalent_Anchoring_of_Thymol_Zein_Nanoparticles_on_Wool_Fibers_via_Polylysine_Cross-linking_for_Durable_Antibacterial_Performance/32147880 CC BY-NC 4.0

oai:figshare.com:article/32147877 2026-05-02T00:00:00Z category_1 category_4 category_8 category_13 category_21 category_46 category_69 category_734 category_873 portal_63 item_type_6 month_year_05_2026

Discovery and Characterization of a Novel Psychrophilic Alginate Lyase Belonging to the New PL7_7 Subfamily Heng Zhang (320479) Ziyan Song (18498857) Jiayi Sun (754336) Yingjie Li (2106778) Lushan Wang (229439) Biophysics Biochemistry Microbiology Genetics Biotechnology Immunology Inorganic Chemistry Biological Sciences not elsewhere classified Chemical Sciences not elsewhere classified structural disulfide bond function studies showed diversity known within disulfide motif appears biochemical analyses revealed 0 – 8 0 ), salt major end products cbms modulate activity novel alginate lyase new pl7_7 subfamily pl7_7 subfamily major component enzymatic activity degrading alginate alginate degradation work broadens vibrio </ study characterizes promising cold product profiles potential applications pl7 family ph 6 linker 2 large number conserved cysteine connecting cbm32 biomedical industries adapted biocatalyst Alginate lyases are crucial for degrading alginate, a major component of brown algal cell walls, yet a large number of alginate lyases remain unclassified. This study characterizes a novel alginate lyase, VzAly7A, from Vibrio sp. B1Z05, assigned to the new PL7_7 subfamily. Biochemical analyses revealed that VzAly7A is psychrophilic (active at 4–30 °C), neutral (pH 6.0–8.0), salt-tolerant, and polyG-preferring, producing unsaturated di-, tri-, and tetrasaccharides as major end products. Domain-function studies showed that CBMs modulate activity and product profiles, and that linker 2, connecting CBM32 to the catalytic domain, is essential for alginate degradation. A conserved cysteine (Cys368) within linker 2 forms a structural disulfide bond with Cys414, which is critical for enzymatic activity. This disulfide motif appears to be unique to the PL7_7 subfamily. This work broadens the diversity known within the PL7 family and provides a promising cold-adapted biocatalyst with potential applications in food, agricultural, and biomedical industries. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acs.jafc.6c01734.s001 https://figshare.com/articles/journal_contribution/Discovery_and_Characterization_of_a_Novel_Psychrophilic_Alginate_Lyase_Belonging_to_the_New_PL7_7_Subfamily/32147877 CC BY-NC 4.0

oai:figshare.com:article/32147849 2026-05-02T00:00:00Z category_4 category_7 category_21 category_39 category_146 category_272 category_873 portal_63 item_type_6 month_year_05_2026

Hydrophobic Carbon with Abundant Defects and Nitrogen Enrichment Effectively Enhances Adsorption of Styrene: Experiments and DFT Calculations Huaxin Xiong (23827514) Jun Liu (42548) Zipei Zhang (7249754) Xiaoqing Liu (196900) Qiang Ren (1565044) Ying Wang (11406) Yongfa Zhang (1298994) Yuqiong Zhao (1743904) Guojie Zhang (146963) Yongliang Ma (2611399) Jingcheng Hao (1404541) Junhua Li (125998) Biochemistry Medicine Biotechnology Ecology Space Science Environmental Sciences not elsewhere classified Chemical Sciences not elsewhere classified volatile organic compounds thereby reducing competition structural characterization demonstrated optimal balance among moisture resistance remains doped porous carbons density functional theory comprehensive elemental analysis specific surface area n – 6 highest nitrogen content 280 ° c styrene adsorption capacity surface chemistry derived n − 18 vocs ). supercritical treatment study offers still able simultaneously increasing sample prepared retain 80 relative humidity pretreatment temperature pore architecture pncs ). hydrophobic carbon humid conditions high selectivity g ). fabricate lignite effective removal doping strategy critical challenge cost adsorbents calculations indicate adsorption energy abundant defects 82 %). 58 kj 43 kj 280 exhibits 1329 mg Developing low-cost adsorbents with high selectivity and moisture resistance remains a critical challenge for the effective removal of volatile organic compounds (VOCs). Herein, a synergistic methanol supercritical pretreatment and nitrogen-doping strategy was proposed to fabricate lignite-derived N-doped porous carbons (PNCs). Comprehensive elemental analysis and structural characterization demonstrated that the sample prepared at a pretreatment temperature of 280 °C (PNC-280) achieved an optimal balance among the degree of defect, pore architecture, surface chemistry, and hydrophobicity. After supercritical treatment, PNC-280 exhibits a specific surface area of 1033 m2/g and the highest nitrogen content (5.82%). Even at 50% relative humidity, PNC-280 was still able to retain 80% of its styrene adsorption capacity (1329 mg/g). Density functional theory (DFT) calculations indicate that the adsorption energy of N–6 for styrene is −18.58 kJ/mol, while that for water is −5.43 kJ/mol, thereby reducing competition from water vapor and simultaneously increasing the affinity for styrene. This study offers a solution for enhancing the adsorption of styrene under humid conditions. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acs.langmuir.6c01196.s001 https://figshare.com/articles/journal_contribution/Hydrophobic_Carbon_with_Abundant_Defects_and_Nitrogen_Enrichment_Effectively_Enhances_Adsorption_of_Styrene_Experiments_and_DFT_Calculations/32147849 CC BY-NC 4.0

oai:figshare.com:article/32147846 2026-05-02T00:00:00Z category_1 category_14 category_19 category_21 category_39 category_734 category_873 category_915 portal_63 item_type_6 month_year_05_2026

Reversible Photoluminescence Switching in Manganese Hybrids Induced by Water-Chloride Exchange Wei Liang (26352) Jiawei Lin (4970902) Tian-Yi Li (1694851) Ruonan Yao (14268269) Yibo Cui (18691765) Zhongnan Guo (4970899) Jing Zhao (21160) Quanlin Liu (1412533) Wenxia Yuan (3778042) Biophysics Molecular Biology Pharmacology Biotechnology Ecology Biological Sciences not elsewhere classified Chemical Sciences not elsewhere classified Physical Sciences not elsewhere classified whose response mechanism theoretical analyses reveal sup >–</ sup responsive luminescent materials hybrid manganese chlorides alternative chromic mechanism manganese hybrids induced based halides provide reversible photoluminescence switching ligand exchange induced 8 </ sub 2 </ sub related pl switching chromic behavior induced chloride exchange mn 4 < sub )< sub generally based isolated mn two zero triggered via tetrahedral (< reported herein red emission organic cations octahedral configuration new platform hydrochloric acid emerging stimuli diaminopyrimidinium cations developing intelligent cyan emission anticounterfeiting patterns Mn-based hybrids are emerging stimuli-responsive luminescent materials, whose response mechanism is generally based on chromic behavior induced by a transition from octahedral (Oh) to tetrahedral (Td) coordination. Upon a reversible phase transition driven by H2O/Cl– ligand exchange in an octahedral configuration (Oh to Oh), an alternative chromic mechanism is reported herein. Two zero-dimensional (0D) hybrid manganese chlorides, (C4H8N4)2[MnCl4(H2O)2]·2Cl (Mn-c) and (C4H8N4)2MnCl6 (Mn-r), were synthesized, featuring 4,6-diaminopyrimidinium cations and isolated Mn-centered octahedral motifs. Structural, spectroscopic, and theoretical analyses reveal that the cyan emission of Mn-c originates from the organic cations, whereas the red emission of Mn-r stems from Mn-centered d-d transitions. Interconversion between Mn-c and Mn-r can be triggered via the H2O/Cl– ligand exchange induced by heating (>108 °C) or by soaking in hydrochloric acid, resulting in a reversible phase transition and switchable photoluminescence (PL) behavior. Leveraging this reversible chromism, anticounterfeiting patterns were fabricated. This novel Oh-Oh phase transition and the related PL switching of 0D hybrid Mn-based halides provide a new platform for developing intelligent, multicolor-responsive luminescent materials. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acs.inorgchem.6c00723.s001 https://figshare.com/articles/journal_contribution/Reversible_Photoluminescence_Switching_in_Manganese_Hybrids_Induced_by_Water-Chloride_Exchange/32147846 CC BY-NC 4.0

oai:figshare.com:article/32147843 2026-05-02T00:00:00Z category_1 category_8 category_21 category_734 category_873 category_915 portal_63 item_type_6 month_year_05_2026

Hydrogen-Bonded Supramolecular Cholesteric Liquid Crystal Polymer Films with Reversible Stimuli Responsiveness Junjie Ren (10993514) Guancheng Lin (21798128) Hairong Yu (6387938) Xingbin Lv (10143197) Changjing Cheng (6387941) Ting Liang (506921) Biophysics Microbiology Biotechnology Biological Sciences not elsewhere classified Chemical Sciences not elsewhere classified Physical Sciences not elsewhere classified visible color changes responsive photonic crystals outstanding regeneration performance liquid crystal monomers hold great promise color display devices tunable structural colors initial structural color demonstrating excellent reusability structural stability excellent sensor water immersion systematically investigated spcs ), spc film reversible response reversible nature reversible dynamics results reveal resulting film response speed response rate reflection wavelength polymerization time moderately shortening innovative packaging information encryption film exhibits external stimuli environmental stimuli environmental monitoring enabling fast effectively enhance chiral dopants biomedical diagnostics Stimuli-responsive photonic crystals (SPCs), with their tunable structural colors and responsiveness to external stimuli, hold great promise for applications in anticounterfeiting, information encryption, and color display devices. In this work, we report a high-performance SPC polymer film based on a hydrogen-bonded supramolecular cholesteric liquid crystal system. Due to the dynamic and reversible nature of hydrogen bonding, this film exhibits a rapid, reversible response to environmental stimuli. The control of the initial structural color was achieved by adjusting the ratio of liquid crystal monomers to chiral dopants. The resulting film can directly convert external signals, such as pH, temperature, and water immersion, into shifts in the reflection wavelength and visible color changes, enabling fast, intuitive visual detection without external power sources. Benefiting from the reversible dynamics of hydrogen bonding, the film maintains robust mechanical strength and structural stability after repeated acid–base cycling, demonstrating excellent reusability. Furthermore, the effects of film thickness, ambient temperature, and polymerization time on the response speed were systematically investigated. The results reveal that reducing the thickness, increasing the ambient temperature, and moderately shortening the polymerization time can effectively enhance the response rate. Combining multicolor reversible transitions, high stability, and outstanding regeneration performance, such an SPC film can serve as an excellent sensor and offers broad application prospects in environmental monitoring, innovative packaging, and biomedical diagnostics. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acsapm.6c00726.s001 https://figshare.com/articles/journal_contribution/Hydrogen-Bonded_Supramolecular_Cholesteric_Liquid_Crystal_Polymer_Films_with_Reversible_Stimuli_Responsiveness/32147843 CC BY-NC 4.0

oai:figshare.com:article/32147840 2026-05-02T00:00:00Z category_13 category_21 category_39 category_133 category_146 category_272 category_734 category_873 portal_63 item_type_6 month_year_05_2026

Boosting Electricity Generation from Ambient Water Evaporation Via Natural Wood-Based Porous Generator Zhenghong Liu (2846237) Kuan Wang (571816) Qing Chang (93814) Ning Li (45258) Chaorui Xue (1680697) Suping Jia (1758274) Jinlong Yang (516710) Shengliang Hu (1902016) Genetics Biotechnology Ecology Plant Biology Space Science Environmental Sciences not elsewhere classified Biological Sciences not elsewhere classified Chemical Sciences not elsewhere classified salt resistance capacity larger surface area induced electricity generators induced electricity generation holding great promise boosting electricity generation based porous generators attracting increasing interest ambient water evaporation delignified balsa wood sustainable water evaporation developed pdw generator green balsa wood obtained pva hydrogel natural wood sustainable energy pdw ). vinyl alcohol synergistic effect study provides simple fabrication produce renewable practical applications nacl solution multiscale channels improved hydrophilicity favorable characteristics dw ). developing cost containing groups complicated synthesis coating poly coated dw Water evaporation-induced electricity generation as a green and effective technique to produce renewable and sustainable energy is attracting increasing interest. Natural wood-based porous generators are promising but remain some challenges including limited output performance and complicated synthesis. Here, a green balsa wood (BW)-based electricity generator from ambient water evaporation was constructed by simple fabrication of coating poly(vinyl alcohol) (PVA) hydrogel on delignified balsa wood (DW). The synergistic effect of delignification and PVA hydrogel coating produced more multiscale channels and oxygen-containing groups, which led to a larger surface area, improved hydrophilicity, and higher surface charge density of the obtained PVA hydrogel-coated DW (PDW). Benefiting from these favorable characteristics, the developed PDW generator (PDWG) showed efficient power generation performance under ambient water evaporation. Besides, it was found that the performance of PDWG can be enhanced in a NaCl solution. Moreover, the PDWG possessed good stability and salt resistance capacity, holding great promise for practical applications. This study provides some implications for developing cost-effective and sustainable water evaporation-induced electricity generators with high performance. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acsapm.6c00619.s001 https://figshare.com/articles/journal_contribution/Boosting_Electricity_Generation_from_Ambient_Water_Evaporation_Via_Natural_Wood-Based_Porous_Generator/32147840 CC BY-NC 4.0

oai:figshare.com:article/32147791 2026-05-02T00:00:00Z category_4 category_7 category_12 category_14 category_19 category_21 category_61 category_64 category_133 category_135 category_734 category_873 portal_63 item_type_6 month_year_05_2026

Size-Tunable Positively Charged Methacrylated Gelatin/Chitosan Composite Hydrogel Microspheres Promote Angiogenesis Yu Song (316955) Zhiqi Lou (23827456) Chang Luo (404131) Danyang Song (15225485) Jiekun Sun (23827459) Qiu Zhao (8904482) Miao Xu (130624) Ziyu Liu (2566468) Zhonghua Liu (148625) Tingsheng Yan (3901732) Biochemistry Medicine Cell Biology Molecular Biology Pharmacology Biotechnology Developmental Biology Cancer Plant Biology Computational Biology Biological Sciences not elsewhere classified Chemical Sciences not elsewhere classified treating tissue defects mediated cellular interactions induced radical polymerization functional tissue regeneration excellent elastic recovery enhancing cellular affinity dependent mechanical feedback blood flow recovery based modification method csma incorporation imparted tunable porous gelma positive surface charge csma composite microspheres hydrogel microspheres uv light tube formation synthesized via subcutaneous implantation significantly promoted regenerative microenvironment regenerative medicine promote vascularization organ damage optimized formulation ischemic diseases huvecs ), gelatin methacryloyl fundamental prerequisite favorable pro effective strategy controllable degradability chitosan methacryloyl Angiogenesis is a fundamental prerequisite for functional tissue regeneration, and biomaterials that drive endogenous vascularization hold immense translational potential for treating tissue defects, organ damage, and ischemic diseases. Herein, gelatin methacryloyl (GelMA) and chitosan methacryloyl (CSMA) were synthesized via a copolymerization-based modification method. Hydrogel microspheres were prepared by emulsification, followed by cross-linking through photoinitiator-induced radical polymerization under UV light. Combined with freeze-drying, size-tunable porous GelMA/CSMA composite microspheres (G/CMS) were fabricated. The as-prepared G/CMS establish a favorable pro-regenerative microenvironment by integrating size-dependent mechanical feedback and charge-mediated cellular interactions. Specifically, CSMA incorporation imparted a positive surface charge, enhancing cellular affinity, while smaller diameters amplified mechanical stimuli promoting adhesion via mechanotransduction. In vitro, the optimized formulation (G/CMS-B) significantly promoted the proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs), and upregulated key angiogenic genes (VEGF, ANG, KDR) without exogenous growth factors. In vivo, subcutaneous implantation and hindlimb ischemia models confirmed accelerated neovascularization and blood flow recovery. The developed G/CMS exhibited excellent biocompatibility, controllable degradability, injectability, and excellent elastic recovery. This synergistic platform effectively modulates physicochemical cues to promote vascularization, offering a promising, cost-effective strategy for regenerative medicine. 2026-05-02T00:00:00Z Text Journal contribution 10.1021/acsami.6c03715.s001 https://figshare.com/articles/journal_contribution/Size-Tunable_Positively_Charged_Methacrylated_Gelatin_Chitosan_Composite_Hydrogel_Microspheres_Promote_Angiogenesis/32147791 CC BY-NC 4.0

oai:figshare.com:article/32147748 2026-05-02T00:00:00Z category_8 category_13 category_14 category_24 category_135 category_146 portal_63 item_type_3 month_year_05_2026

Cross-Strain Transferability of CRISPRi Systems and Design Rules from Laboratory to Clinical Escherichia coli Strains Hyerim Ban (23827422) Stephen N. Rondthaler (17774803) Matthew Lebovich (1529167) Marcos A. Lora (23827425) Brandon Ugbesia (23827428) Lauren B. Andrews (12911000) Microbiology Genetics Molecular Biology Evolutionary Biology Computational Biology Space Science two dcas12a variants showed high variation probiotic nissle 1917 present important limitations findings provide insight specific transcriptional silencing uropathogenic umn026 ). notably poorer repression specific design considerations strain using sets three crispri systems crispri genetic system crispri genetic design three nonmodel strains multiple clinical strains targeted gene repression crispri systems crispri system specific effects nonmodel strains simple design design rules clinical isolates repression efficiency fold repression coli strains well understood versatile approach uropathogenic cft073 targeting along strain transferability results demonstrate providing guidelines one dcas9 many organisms guide rnas diverse applications dcas protein criteria used assayed cytotoxicity 12 strain CRISPR interference (CRISPRi) has emerged as a versatile approach for targeted gene repression in many organisms, including microbes and bacteria, due to the simple design of sequence-specific transcriptional silencing of gene expression. However, the strain-specific effects on repression efficiency and the host when translating a CRISPRi system from a laboratory strain to nonmodel strains are not well understood, yet they can present important limitations to its use. Here, we investigated the repression efficiency and toxicity of three CRISPRi systems (one dCas9 and two dCas12a variants) across four different Escherichia coli strains, including a laboratory K-12 strain (MG1655) and three nonmodel strains that are clinical isolates (probiotic Nissle 1917, uropathogenic CFT073, and uropathogenic UMN026). We evaluated the repression in each strain using sets of guide RNAs (gRNAs) targeting along the gene sequence and assayed cytotoxicity of expressing each dCas protein. Growth toxicity from expression of the different dCas proteins notably differed and showed high variation between some host strains. We also observed variable repression among the strains and notably poorer repression in multiple clinical strains. Therefore, we developed a dual gRNA CRISPRi system for enhanced gene silencing among the strains, which achieved up to 824-fold repression in CFT073. The results demonstrate that strain-specific design considerations can arise when a CRISPRi genetic system is transferred to a closely related bacterial strain. These findings provide insight into the relationships between criteria used for CRISPRi genetic design and in vivo activity across nonmodel E. coli strains, providing guidelines for diverse applications of these tools. 2026-05-02T00:00:00Z Dataset Dataset 10.1021/acssynbio.6c00075.s004 https://figshare.com/articles/dataset/Cross-Strain_Transferability_of_CRISPRi_Systems_and_Design_Rules_from_Laboratory_to_Clinical_Escherichia_coli_Strains/32147748 CC BY-NC 4.0

oai:figshare.com:article/32147745 2026-05-02T00:00:00Z category_8 category_13 category_14 category_24 category_135 category_146 portal_63 item_type_3 month_year_05_2026

Cross-Strain Transferability of CRISPRi Systems and Design Rules from Laboratory to Clinical Escherichia coli Strains Hyerim Ban (23827422) Stephen N. Rondthaler (17774803) Matthew Lebovich (1529167) Marcos A. Lora (23827425) Brandon Ugbesia (23827428) Lauren B. Andrews (12911000) Microbiology Genetics Molecular Biology Evolutionary Biology Computational Biology Space Science two dcas12a variants showed high variation probiotic nissle 1917 present important limitations findings provide insight specific transcriptional silencing uropathogenic umn026 ). notably poorer repression specific design considerations strain using sets three crispri systems crispri genetic system crispri genetic design three nonmodel strains multiple clinical strains targeted gene repression crispri systems crispri system specific effects nonmodel strains simple design design rules clinical isolates repression efficiency fold repression coli strains well understood versatile approach uropathogenic cft073 targeting along strain transferability results demonstrate providing guidelines one dcas9 many organisms guide rnas diverse applications dcas protein criteria used assayed cytotoxicity 12 strain CRISPR interference (CRISPRi) has emerged as a versatile approach for targeted gene repression in many organisms, including microbes and bacteria, due to the simple design of sequence-specific transcriptional silencing of gene expression. However, the strain-specific effects on repression efficiency and the host when translating a CRISPRi system from a laboratory strain to nonmodel strains are not well understood, yet they can present important limitations to its use. Here, we investigated the repression efficiency and toxicity of three CRISPRi systems (one dCas9 and two dCas12a variants) across four different Escherichia coli strains, including a laboratory K-12 strain (MG1655) and three nonmodel strains that are clinical isolates (probiotic Nissle 1917, uropathogenic CFT073, and uropathogenic UMN026). We evaluated the repression in each strain using sets of guide RNAs (gRNAs) targeting along the gene sequence and assayed cytotoxicity of expressing each dCas protein. Growth toxicity from expression of the different dCas proteins notably differed and showed high variation between some host strains. We also observed variable repression among the strains and notably poorer repression in multiple clinical strains. Therefore, we developed a dual gRNA CRISPRi system for enhanced gene silencing among the strains, which achieved up to 824-fold repression in CFT073. The results demonstrate that strain-specific design considerations can arise when a CRISPRi genetic system is transferred to a closely related bacterial strain. These findings provide insight into the relationships between criteria used for CRISPRi genetic design and in vivo activity across nonmodel E. coli strains, providing guidelines for diverse applications of these tools. 2026-05-02T00:00:00Z Dataset Dataset 10.1021/acssynbio.6c00075.s003 https://figshare.com/articles/dataset/Cross-Strain_Transferability_of_CRISPRi_Systems_and_Design_Rules_from_Laboratory_to_Clinical_Escherichia_coli_Strains/32147745 CC BY-NC 4.0

oai:figshare.com:article/32147742 2026-05-02T00:00:00Z category_8 category_13 category_14 category_24 category_135 category_146 portal_63 item_type_3 month_year_05_2026

Cross-Strain Transferability of CRISPRi Systems and Design Rules from Laboratory to Clinical Escherichia coli Strains Hyerim Ban (23827422) Stephen N. Rondthaler (17774803) Matthew Lebovich (1529167) Marcos A. Lora (23827425) Brandon Ugbesia (23827428) Lauren B. Andrews (12911000) Microbiology Genetics Molecular Biology Evolutionary Biology Computational Biology Space Science two dcas12a variants showed high variation probiotic nissle 1917 present important limitations findings provide insight specific transcriptional silencing uropathogenic umn026 ). notably poorer repression specific design considerations strain using sets three crispri systems crispri genetic system crispri genetic design three nonmodel strains multiple clinical strains targeted gene repression crispri systems crispri system specific effects nonmodel strains simple design design rules clinical isolates repression efficiency fold repression coli strains well understood versatile approach uropathogenic cft073 targeting along strain transferability results demonstrate providing guidelines one dcas9 many organisms guide rnas diverse applications dcas protein criteria used assayed cytotoxicity 12 strain CRISPR interference (CRISPRi) has emerged as a versatile approach for targeted gene repression in many organisms, including microbes and bacteria, due to the simple design of sequence-specific transcriptional silencing of gene expression. However, the strain-specific effects on repression efficiency and the host when translating a CRISPRi system from a laboratory strain to nonmodel strains are not well understood, yet they can present important limitations to its use. Here, we investigated the repression efficiency and toxicity of three CRISPRi systems (one dCas9 and two dCas12a variants) across four different Escherichia coli strains, including a laboratory K-12 strain (MG1655) and three nonmodel strains that are clinical isolates (probiotic Nissle 1917, uropathogenic CFT073, and uropathogenic UMN026). We evaluated the repression in each strain using sets of guide RNAs (gRNAs) targeting along the gene sequence and assayed cytotoxicity of expressing each dCas protein. Growth toxicity from expression of the different dCas proteins notably differed and showed high variation between some host strains. We also observed variable repression among the strains and notably poorer repression in multiple clinical strains. Therefore, we developed a dual gRNA CRISPRi system for enhanced gene silencing among the strains, which achieved up to 824-fold repression in CFT073. The results demonstrate that strain-specific design considerations can arise when a CRISPRi genetic system is transferred to a closely related bacterial strain. These findings provide insight into the relationships between criteria used for CRISPRi genetic design and in vivo activity across nonmodel E. coli strains, providing guidelines for diverse applications of these tools. 2026-05-02T00:00:00Z Dataset Dataset 10.1021/acssynbio.6c00075.s002 https://figshare.com/articles/dataset/Cross-Strain_Transferability_of_CRISPRi_Systems_and_Design_Rules_from_Laboratory_to_Clinical_Escherichia_coli_Strains/32147742 CC BY-NC 4.0 eyJtZXRhZGF0YVByZWZpeCI6ICJvYWlfZGMiLCAidmVyYiI6ICJMaXN0UmVjb3JkcyIsICJvYWlfaGFuZGxlciI6IHRydWUsICJpbnN0aXR1dGlvbl9pZCI6IDYzLCAicGFnZSI6IDIsICJ4Q3Vyc29yIjogImV5SmhiR2NpT2lKSVV6VXhNaUo5Lklsc3hOemMzTnpReE5qazRNREF3TENBek1qRTBOemMwTWwwaS5ibGRQVnBuV2w5M2ZKMkkzUlVUWU1FNFFUUG9vRmRjMEhFM0J2Y1JoYVR3ZV80dFdWZ0x2STdxOEVqcGd0U01YOThybnJDUjFMM0RNMTgyWW9XYjFVdyIsICJleHBpcmF0aW9uIjogIjIwMjYtMDUtMDMgMDk6MjA6NTIifQ==