New biotechnology最新文献_第2页

Screening and bioreactor cultivation of wild-type lactic acid bacteria for high purity D-lactic acid production 高纯度d-乳酸野生型乳酸菌的筛选及生物反应器培养

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-08-20 DOI: 10.1016/j.nbt.2025.08.005

Laura Troiani , Alessia Levante , Hannes Russmayer , Hans Marx , Erasmo Neviani , Valentina Bernini , Camilla Lazzi , Michael Sauer

{"title":"Screening and bioreactor cultivation of wild-type lactic acid bacteria for high purity D-lactic acid production","authors":"Laura Troiani , Alessia Levante , Hannes Russmayer , Hans Marx , Erasmo Neviani , Valentina Bernini , Camilla Lazzi , Michael Sauer","doi":"10.1016/j.nbt.2025.08.005","DOIUrl":"10.1016/j.nbt.2025.08.005","url":null,"abstract":"<div><div>Biodegradable polymers are the green alternative to conventional oil-based plastics and have a key role in the achievement of the Sustainable Development Goals of Agenda 2030. Nowadays, polylactic acid (PLA) is one of the most common bioplastics present in the global market. Optically pure D-lactic acid (D-LA) is a valuable monomer for the production of high-performance PLA materials. This study aims to select wild-type lactic acid bacteria capable of producing D-LA with high optical purity and to define an optimized fermentation process in bioreactors to maximize production. A total of 150 LAB strains from the University of Parma Culture Collection (UPCCO) were screened using enzymatic assays and HPLC analysis. Among them, <em>Lactobacillus delbrueckii</em> UPCCO 2214 and <em>Leuconostoc citreum</em> UPCCO 4516 were selected for fed-batch cultivations for their high D-LA production and purity (>97 %). After optimization of fermentation parameters, <em>L. delbrueckii</em> UPCCO 2214 was found to be the most efficient strain for D-LA production, with a yield of 0.74 g/g and a volumetric productivity of 0.96 g/L/h, outperforming <em>L. citreum</em> UPCCO 4516. Bioreactors cultivation has helped to understand microbial production and boost their potentialities. This work supports further investigations and improvements in D-LA production processes to advance the field of biomaterials with benefits for both industry and the environment.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 48-55"},"PeriodicalIF":4.9,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144894877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Lactobacillus salivarius HHuMin-U attenuates vulvovaginal candidiasis via vaginal epithelial immune enhancement mediated by NF-κB activation 唾液乳杆菌HHuMin-U通过NF-κB活化介导的阴道上皮免疫增强，减轻外阴阴道念珠菌病

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-08-19 DOI: 10.1016/j.nbt.2025.08.003

Juwon Choi , Jay-Young Jo , Jisu Lee , Jo Eun Son , Sun Young Kim , Hye Eun Lee , Yeong-Je Seong , Keon Heo , Yongbaek Kim , Myeong Soo Park , Sanguine Byun

{"title":"Lactobacillus salivarius HHuMin-U attenuates vulvovaginal candidiasis via vaginal epithelial immune enhancement mediated by NF-κB activation","authors":"Juwon Choi , Jay-Young Jo , Jisu Lee , Jo Eun Son , Sun Young Kim , Hye Eun Lee , Yeong-Je Seong , Keon Heo , Yongbaek Kim , Myeong Soo Park , Sanguine Byun","doi":"10.1016/j.nbt.2025.08.003","DOIUrl":"10.1016/j.nbt.2025.08.003","url":null,"abstract":"<div><div>Vulvovaginal candidiasis (VVC), primarily caused by <em>Candida albicans</em> infection, is one of the most common diseases among women and leads to various symptoms that adversely impact quality of life. VVC is conventionally treated with antifungal agents, but the high recurrence rates and the risk of inducing vaginal microbiome dysbiosis pose major concerns in effective treatment. Probiotics with immune-enhancing properties can be an effective strategy by strengthening mucosal immunity and reducing susceptibility to infection. Herein, this study investigates the therapeutic potential of <em>Lactobacillus salivarius</em> HHuMin-U (HMU) as a probiotic agent for treating VVC. Phenotype screening identified HMU as a top candidate with antifungal activity against <em>C. albicans</em>. HMU significantly upregulated immunomodulatory factors such as antimicrobial peptides, cytokines, and chemokines in human vaginal epithelial cells, which can strengthen the antifungal immune system. In an animal study, administration of HMU in a mouse VVC model significantly decreased the fungal burden and protected the mice from vaginal infection. Additionally, cellular infection models revealed that HMU reduced fungal adhesion and the cytolytic activity of <em>C. albicans</em>, while conditioned media from HMU-treated epithelial cells inhibited fungal growth. Transcriptomic analysis revealed that HMU treatment enriched gene sets related to epithelial barrier integrity, innate immune responses mediated by epithelial cells, and immune cell regulation. Mechanistically, the NF-κB pathway emerged as a key mediator of these responses. Collectively, HMU inhibits VVC by enhancing epithelial immunity and thus could be considered as a probiotic agent for the prevention and treatment of VVC.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 36-47"},"PeriodicalIF":4.9,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889218","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterization and orthogonality assessment of two quorum sensing systems for synthetic biology applications 合成生物学应用中两种群体感应系统的特性和正交性评价

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-08-18 DOI: 10.1016/j.nbt.2025.08.001

Jasmine De Baets, Brecht De Paepe, Marjan De Mey

{"title":"Characterization and orthogonality assessment of two quorum sensing systems for synthetic biology applications","authors":"Jasmine De Baets, Brecht De Paepe, Marjan De Mey","doi":"10.1016/j.nbt.2025.08.001","DOIUrl":"10.1016/j.nbt.2025.08.001","url":null,"abstract":"<div><div>Quorum sensing systems have a broad range of applications within the field of synthetic biology. However, a bottleneck is the optimization and tuning of these systems due to the lack of standardization and complete characterization. In this research, two quorum sensing systems, namely the LasI/LasR and the EsaI/EsaR system, were fully characterized in the model host organism <em>Escherichia coli</em>. Furthermore, insight was gained in the interplay between the various parts of these systems. To further expand the range of possibilities with these quorum sensing systems, the orthogonality of the two systems was assessed to allow simultaneous use within the same cell without interfering crosstalk. This assessment was performed on three levels: promoter, signal and synthase crosstalk. It was demonstrated that LasR is able to interact with the promoter of the EsaI/EsaR system, albeit to a low extent. Additionally, LasR was able to respond to the autoinducers produced by EsaI. To solve the promoter crosstalk, a nucleotide change was introduced into the binding site of EsaR within the promoter region. Additionally, LasR mutants were created rationally and screened for decreased response to EsaI while retaining functionality. The best performing mutant, LasR(P117S), was further characterized. In conclusion, we have further unlocked the potential of quorum sensing systems for synthetic biology applications by obtaining two functional, characterized and orthogonal quorum sensing systems.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 20-35"},"PeriodicalIF":4.9,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144886643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Public perception of new plant breeding techniques for sustainable production of feed and food in the Czech Republic 公众对捷克共和国饲料和食品可持续生产的新植物育种技术的看法

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-08-13 DOI: 10.1016/j.nbt.2025.08.002

Naděžda Čadová , Jaroslav Doležel , Ivo Frébort

{"title":"Public perception of new plant breeding techniques for sustainable production of feed and food in the Czech Republic","authors":"Naděžda Čadová , Jaroslav Doležel , Ivo Frébort","doi":"10.1016/j.nbt.2025.08.002","DOIUrl":"10.1016/j.nbt.2025.08.002","url":null,"abstract":"<div><div>A survey was conducted among 1676 Czech citizens in the autumn of 2024 to explore their attitudes toward different methods of crop improvement, which are based on the modification of genetic information. The study focused on three techniques: classical mutagenesis, insertion of foreign genetic material, and genome editing. The findings revealed that a significant part of the general public was unfamiliar with these technologies and their legal status. A majority of respondents mistakenly believed that currently permitted mutation breeding using radiation or chemical mutagens is illegal, while conversely assuming that techniques strictly regulated in Europe, such as transgenesis and genome editing, are allowed. Despite using different survey methods and question wording, the study pointed to a modest positive shift in public attitudes toward new genetic technologies compared to earlier surveys. For example, support for legalising genome editing rose from 22 % in 2019 to 47.6 % in 2024, and the proportion of respondents who would buy food containing genetically modified ingredients increased from 35 % to 47.4 %. Acceptance of these methods varied across demographic groups, with younger, more educated, and male respondents showing higher levels of support. While many acknowledge potential benefits such as reduced pesticide use, concerns about safety, ethics, and environmental impact remain. In general, the Czech public demonstrates cautious optimism, underscoring the importance of transparency and clear product labelling.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 13-19"},"PeriodicalIF":4.9,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144829657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Viability assessment and biomass valorization of recombinant Komagataella phaffii 重组法菲Komagataella phaffii的活力评价与生物量增值。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-08-05 DOI: 10.1016/j.nbt.2025.07.005

Sarah Gangl , Maik Hilgarth , Christian Spielhaupter , Frank Schneider , Christiane Herrmann , Christina Recher , Tobias Baumgartner , Martina Geier , Birgit Wassermann , Anton Glieder

{"title":"Viability assessment and biomass valorization of recombinant Komagataella phaffii","authors":"Sarah Gangl , Maik Hilgarth , Christian Spielhaupter , Frank Schneider , Christiane Herrmann , Christina Recher , Tobias Baumgartner , Martina Geier , Birgit Wassermann , Anton Glieder","doi":"10.1016/j.nbt.2025.07.005","DOIUrl":"10.1016/j.nbt.2025.07.005","url":null,"abstract":"<div><div><em>Komagataella phaffii</em> has become a key player in biotechnology used for the production of heterologous proteins with numerous GRAS notices and QPS status released in the past years. While most of these descriptions and scientific studies focus on strain engineering, products and cultivation processes, this paper provides fundamental data about the general viability of <em>K. phaffii</em> after post-production biomass treatments and co-incubations with non-sterile soil and water samples. A comparison of different <em>K. phaffii</em> strains uncovered a high thermal sensitivity for all tested strains, which was further enhanced under alkaline conditions. A rapid reduction in the viable cell number started at approx. 52 °C and terminated in a full loss of 10<sup>9</sup> viable cells per milliliter within seconds at 70 °C. Survival rates of untreated non-GMO and GMO <em>K. phaffii</em> strains in water- and soil-slurry-microcosms demonstrated that the number of viable <em>K. phaffii</em> cells decreased rapidly and comparably to a <em>Saccharomyces cerevisiae</em> reference. No mating events were detected under these environmental conditions. These findings supported assumptions that <em>K. phaffii</em> is highly specialized and not a ubiquitous microorganism as described for other yeasts. Finally, intrigued by the low survival outside optimal laboratory conditions, <em>K. phaffii</em> biomass was evaluated in mesophilic anaerobic digestion processes for energy production. Enhanced methane yields and a concomitant loss of viable <em>K. phaffii</em> cells were demonstrated.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 1-12"},"PeriodicalIF":4.9,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144775816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Creation of colorless transparent tilapia using CRISPR/Cas9 mediated multi-gene mutation 利用CRISPR/Cas9介导的多基因突变培育无色透明罗非鱼。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-07-31 DOI: 10.1016/j.nbt.2025.07.009

Guangyuan Liang , Baoyue Lu , Shengfei Dai, Minghui Li, Jiawen Yao, Hao Liu, Xiayue Liu, Xingyong Liu, Deshou Wang

{"title":"Creation of colorless transparent tilapia using CRISPR/Cas9 mediated multi-gene mutation","authors":"Guangyuan Liang , Baoyue Lu , Shengfei Dai, Minghui Li, Jiawen Yao, Hao Liu, Xiayue Liu, Xingyong Liu, Deshou Wang","doi":"10.1016/j.nbt.2025.07.009","DOIUrl":"10.1016/j.nbt.2025.07.009","url":null,"abstract":"<div><div>Transparent mutant fish have been obtained from small-sized fish (medaka, zebrafish, and killifish), and are often used as experimental models in biological and medical research. However, transparent fish have never been created in medium- or large-sized fish through gene editing. In this study, mutants without xanthophores, erythrophores and pigmented melanophores were first obtained by crossing to form <em>tyrb;csf1ra</em> double mutant tilapia. Subsequently, single mutants lacking reflective platelets without visible iridophores were obtained by mutation of <em>pnp4a</em> and <em>tfec</em> using CRISPR/Cas9 gene editing, the recovery of iridophores were observed in <em>pnp4a</em> mutants but not in <em>tfec</em> mutants in the later stage. In addition, we also found that <em>tfec</em> mutation led to a decrease in number of melanophores and diameter of melanophores/erythrophores. Finally, transparent triple mutant homozygotes without visible pigment cells were obtained by triple mutation of <em>tyrb</em>;<em>csf1ra</em>;<em>pnp4a</em> (named <em>amber</em>) and <em>tyrb</em>;<em>csf1ra</em>;<em>tfec</em> (named <em>ruby</em>), through crossing of double and single mutants. The two mutant lines were no longer transparent at 60 dpf (days post fertilization) and 120 dpf, respectively, due to the recovery of iridophores in the former, and thickening of the body wall in the latter. In summary, this study created two transparent lines without visible pigment cells through aggregation of multiple gene mutation and crossing. The transparent fish is suitable for in vivo imaging, and is currently the only medium-sized transparent fish obtained through gene editing.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 163-176"},"PeriodicalIF":4.9,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144768811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biology-aware machine learning for culture medium optimization 用于培养基优化的生物感知机器学习

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-07-25 DOI: 10.1016/j.nbt.2025.07.006

Takamasa Hashizume , Bei-Wen Ying

引用次数: 0

Using control bias to identify initial targets for bioproduction improvement 利用控制偏差确定生物生产改进的初始目标。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-07-24 DOI: 10.1016/j.nbt.2025.07.008

Michael Binns , Pedro de Atauri , Marta Cascante , Constantinos Theodoropoulos

{"title":"Using control bias to identify initial targets for bioproduction improvement","authors":"Michael Binns , Pedro de Atauri , Marta Cascante , Constantinos Theodoropoulos","doi":"10.1016/j.nbt.2025.07.008","DOIUrl":"10.1016/j.nbt.2025.07.008","url":null,"abstract":"<div><div>Sensitivity analysis of bioprocess metabolic reaction networks analysis allows the prediction of system parameters such as those associated with the enzyme activity of certain reaction steps which significantly affect the overall production. However, uncertainties in kinetic rate expressions and in the resulting steady-state flux distributions limit the accuracy of these predictions. Starting from minimal information (reaction stoichiometry, and external fluxes in/out of the system and potentially identification of steps at equilibrium) a new preliminary method is proposed using sampling of elasticities and metabolic fluxes to calculate the control bias. The calculated control bias identifies steps which are likely to have positive control, negative control or negligible/uncertain control. This is intended to give initial guidance before further detailed investigation is carried out, identifying targets for any organism to enhance production of valuable chemicals. As a case study, this methodology is applied to succinic acid bioproduction using <em>Actinobacillus succinogenes</em> and analysis successfully reveals the reaction steps having the greatest positive and negative influence on biosuccinic acid production.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 130-140"},"PeriodicalIF":4.9,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144718160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Future food court in an urban shopping center can supply feedstocks for microbial protein production 未来城市购物中心的美食广场可以为微生物蛋白的生产提供原料。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-07-23 DOI: 10.1016/j.nbt.2025.07.007

Terhi Viinikanoja , Eevi Haajanen , Maria Pajumo, Amanda Lillberg, Emilia Nordlund, Anneli Ritala

{"title":"Future food court in an urban shopping center can supply feedstocks for microbial protein production","authors":"Terhi Viinikanoja , Eevi Haajanen , Maria Pajumo, Amanda Lillberg, Emilia Nordlund, Anneli Ritala","doi":"10.1016/j.nbt.2025.07.007","DOIUrl":"10.1016/j.nbt.2025.07.007","url":null,"abstract":"<div><div>Food courts in urban shopping centers can be future sources of feedstock for microbial protein production. This study explored whether filamentous fungi <em>Trichoderma reesei</em>, <em>Paecilomyces variotii</em> and <em>Rhizopus oligosporus</em> can be cultivated on growth media prepared from orange peels, soft drink waste-mix, expired bread or spent coffee grounds, all existing and potential by-products of future food courts. Furthermore, artificial urine to describe human urine as a future nitrogen source was investigated. Side stream pretreatments facilitated sugar releases up to 171 g L<sup>−1</sup> (expired bread). Cultivation media were prepared from pretreated side streams with 40 g L<sup>−1</sup> sugar. Flask cultivations supplemented with nitrogen and micronutrient increased fungal biomass production. From the nitrogen sources tested, yeast extract (10 g L<sup>−1</sup>) proved effective in media based on soft drink waste-mix and orange peel juice. For the latter artificial urine (5 g L<sup>−1</sup> urea) worked as well. The effects of these nitrogen supplements, along with pH, on biomass production were further optimized using design of experiment methodology and validated in lab-scale bioreactor cultivations. The bioreactor cultivations of <em>P. variotii</em> and <em>T. reesei</em> with soft drink waste-mix or orange peel juice supplemented with yeast extract (13 – 15 g L<sup>−1</sup>) resulted in good biomass production up to 37.8 g L<sup>−1</sup> and 15 g L<sup>−1</sup> dry weight with total protein contents reaching 25 % and 35 % for <em>P. variotii</em> and <em>T. reesei</em>, respectively. Amino acid analyses revealed balanced essential amino acid distributions for human consumption, confirming that future food court by-products are viable substrates for sustainable fungal protein production in a circular economy.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 152-162"},"PeriodicalIF":4.9,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144718159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Conditional guide RNA deactivation by mRNA and small molecule triggers in Saccharomyces cerevisiae 酿酒酵母菌mRNA和小分子触发器的条件引导RNA失活。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-07-17 DOI: 10.1016/j.nbt.2025.07.004

Chenggang Xi , Stephen Chiu , William E. Voje , James M. Carothers , Tae Seok Moon

{"title":"Conditional guide RNA deactivation by mRNA and small molecule triggers in Saccharomyces cerevisiae","authors":"Chenggang Xi , Stephen Chiu , William E. Voje , James M. Carothers , Tae Seok Moon","doi":"10.1016/j.nbt.2025.07.004","DOIUrl":"10.1016/j.nbt.2025.07.004","url":null,"abstract":"<div><div>CRISPR interference (CRISPRi) technologies have revolutionized bioengineering by providing precise tools for gene expression modulation, enabling targeted gene perturbation and metabolic pathway optimization. Despite these advances, achieving dynamic control over gene expression by CRISPR-based regulation remains a challenge due to its inherently static nature. Utilizing toehold-mediated strand displacement and ligand-responsive ribozymes (aptazymes), this study introduces switchable guide RNAs (gRNAs) that facilitate tunable gene expression mediated by mRNA or small molecule signals. We demonstrate complete silencing of gRNA via strategically designed 5’ or 3’ extensions that impede the gRNA spacer or the dCas9 handle, with subsequent restoration of function through sequestration or cleavage of the obstructive sequence. The resulting toehold-embedded or aptazyme-embedded gRNAs can be deactivated by specific signals, including two full-length translatable mRNAs and two small molecule triggers, thereby lifting CRISPRi repression on targeted genes. This modular approach allows for gRNA-based biocomputing through multi-layer or multi-input genetic logic gates in <em>Saccharomyces cerevisiae</em>. Offering a versatile strategy for post-CRISPR regulation in response to environmental signals or cellular states, this methodology expands the toolkit in eukaryotic systems for reversible control of gene expression.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 105-118"},"PeriodicalIF":4.5,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144668090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0