mSphere最新文献

{"title":"Overview of the ASM-IISc Symposium on the One Health Approach to AMR in Bengaluru, India.","authors":"Amitesh Anand, Nishad Matange, Samay Pande, Vasundhra Bhandari","doi":"10.1128/msphere.00211-26","DOIUrl":"10.1128/msphere.00211-26","url":null,"abstract":"<p><p>Antimicrobial resistance (AMR) is a systemic global threat, frequently described as a silent pandemic due to the challenges in attributing mortality to drug-resistant infections. Addressing this crisis necessitates a paradigm shift from viewing AMR as a linear consequence of antibiotic exposure to understanding it as an emergent property of ecological systems and metabolic plasticity. The ASM-IISc Symposium on the One Health Approach to AMR brought together a multidisciplinary cohort to deliberate on the drivers and solutions for resistance at the intersection of human, animal, and environmental health. Key thematic deliberations highlighted the role of technology in predicting resistance evolution and identifying novel targets for drug combinations. Furthermore, the symposium underscored the necessity of ecology-driven surveillance, extending genomic monitoring to environmental reservoirs. Ultimately, these findings establish a roadmap toward proactive, predictive, and integrated One Health strategies. Solutions demand mechanistic research and ecosystem-level interventions that bridge molecular insight with global public health strategy.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0021126"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621860/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148795858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cysteine import contributes to hypochlorous acid resistance in <i>Staphylococcus aureus</i>.","authors":"Abigail G Hall, Aliyah J Collins, Anna Sommers, Ryan Singh, Sasmita Panda, Sujata S Chaudhari, Andrew J Monteith, Vinai C Thomas","doi":"10.1128/msphere.00214-26","DOIUrl":"10.1128/msphere.00214-26","url":null,"abstract":"<p><p>Hypochlorous acid (HOCl) is a potent antimicrobial oxidant produced by innate immune cells to eliminate invading bacterial pathogens, including <i>Staphylococcus aureus</i>. HOCl readily oxidizes thiols, amines, and macromolecules in <i>S. aureus</i>. Yet, some bacterial cells within an HOCl-challenged population withstand this oxidative stress and proceed to establish infection. The mechanisms that enable <i>S. aureus</i> to resist the effects of HOCl remain incompletely understood. Here, we show that cysteine uptake is a major determinant of HOCl resistance in <i>S. aureus</i>. Inactivation of the cysteine transporters <i>tcyABC</i> and <i>tcyP</i> resulted in a dose-dependent increase in sensitivity to HOCl. Cysteine uptake through these transporters helped maintain bacillithiol homeostasis and limit oxidative damage to protein thiols during HOCl stress. Importantly, cysteine uptake was required for the survival of <i>S. aureus</i> during interaction with activated human neutrophils that generate HOCl to kill phagocytosed bacteria. These findings reveal that cysteine uptake expands the thiol buffering capacity of <i>S. aureus</i> and functions as a major mechanism for scavenging HOCl, which may contribute to its persistence in the infected host.IMPORTANCE<i>Staphylococcus aureus</i> is a major human pathogen that causes recurrent and antibiotic-recalcitrant infections. Understanding the mechanisms that allow <i>S. aureus</i> to evade key innate immune defenses is therefore of considerable interest for developing effective intervention strategies. Our findings show that cysteine uptake is a critical component of <i>S. aureus</i> defense against hypochlorous acid (HOCl) produced by innate immune cells. Cysteine functions as a reactive sink for HOCl and thereby prevents depletion of both low-molecular-weight thiols and protein thiols in <i>S. aureus</i>.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0021426"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621853/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148808917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Linear DNA expression cassette encapsulated in lipid nanoparticles confers complete protection against influenza A virus in mice.","authors":"Danh C Lai, The N Nguyen, Giao P Trinh, Hiep L X Vu","doi":"10.1128/msphere.00135-26","DOIUrl":"10.1128/msphere.00135-26","url":null,"abstract":"<p><p>DNA-based vaccines have been developed and tested against a range of infectious agents, including influenza A viruses (IAV). Bacterial plasmid DNA is widely used as a vector for delivering genes encoding vaccine immunogens. However, the inclusion of antibiotic resistance genes in conventional plasmid constructs raises safety concerns. Linear DNA expression cassettes produced via PCR that lack antibiotic resistance genes, and bacterial sequences can serve as a safer alternative vector. In this study, we employed a mouse model of IAV to evaluate the protective efficacy of a lipid nanoparticle (LNP)-encapsulated linear DNA expression cassette containing the hemagglutinin gene of IAV H1N1 produced by PCR amplification. For comparison, a circular plasmid DNA containing the same gene was also constructed. The LNP-encapsulated DNA cassette (LNP-L) exhibited similar physicochemical characteristics, such as particle size, polydispersity index, zeta potential, and encapsulation efficiency, compared to the LNP-encapsulated circular plasmid DNA (LNP-P). However, LNP-L produced significantly lower transgene expression than LNP-P in cell culture. Mice immunized with LNP-L developed a delayed onset of antibody response compared to those immunized with LNP-P, but the magnitudes of both humoral and cell-mediated immune responses were comparable between the two groups after booster immunization. Moreover, mice immunized with LNP-L exhibited both Th1 and Th2 responses, similar to those immunized with LNP-P. Importantly, mice immunized with either LNP-L or LNP-P were fully protected upon challenge with a lethal dose of IAV H1N1. These findings suggest that linear DNA expression cassettes encapsulated in LNPs represent a promising strategy for vaccine development.IMPORTANCELipid nanoparticle (LNP)-encapsulated DNA vaccines represent a promising platform for the rapid development of vaccines against infectious pathogens. However, the inclusion of an antibiotic-resistance gene in plasmid DNA vectors used to deliver vaccine immunogens raises potential safety concerns. This study explores the feasibility of using a linear expression cassette produced via PCR amplification as an alternative system for delivering vaccine immunogens. Compared with conventional plasmid DNA, the linear expression cassette contains only essential elements, including a promoter, the gene encoding the vaccine immunogen, and a polyadenylation signal, and can be produced in a cell-free system. Using a mouse model for influenza A virus, we demonstrate that an LNP-encapsulated linear DNA cassette induces humoral and cell-mediated immune responses and protects immunized mice against a lethal influenza virus challenge. Thus, the LNP-encapsulated linear expression cassette may provide a safer approach for rapid responses to infectious diseases in animals.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0013526"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621868/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148892003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aerosol-based disinfection protocol for aircraft: a solution for the conflict between material compatibility and antimicrobial efficacy.","authors":"Sandro Klafack, Jessica Kohs, Karla A Schwenke, Paul Siller, Janina Reißner, Holger Freese, Alina Below, Tim Lübcke, Stefan Mehler, Bernd Albert, Heiko Barth, Sabine Hoelterhoff, Mathias Streitz, Marcell Engel, Jan Schinköthe, Hendrik A Scheinemann, Marc Thanheiser, Uwe Rösler, Jens Teifke, Sven Reiche","doi":"10.1128/msphere.00542-26","DOIUrl":"10.1128/msphere.00542-26","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Every year, billions of passengers, pets, and livestock are transported by air, which increases the risk of a global spread of pathogens like severe acute respiratory syndrome coronavirus 1 and 2, H1N1 influenza, the Ebola virus, and foot-and-mouth disease virus. However, due to its complexity and high safety standards, aircraft decontamination is limited to manual surface disinfection, and an approved method for disinfecting otherwise inaccessible areas is still unavailable. To address this issue, we present the development and validation of an effective room disinfection method utilizing low concentrations of aerosolized peroxyacetic acid (aPAA), stabilized by hydrogen peroxide, known as \"dry fog.\" Therefore, we determined the minimal effective antimicrobial concentration of aPAA for a wide range of agents, including bacteria, viruses, and bacterial endospores. Then, we applied the developed disinfection protocol to a large panel of relevant materials multiple times in the context of the official material compatibility tests (MCTs) requested by the European Union Aviation Safety Agency (EASA) to demonstrate that its application will not cause a negative effect on the aircraft airworthiness. After successfully passing these complex material compatibility tests, we finally validated the disinfection protocol within a narrow-body aircraft A320 for its antimicrobial efficacy by using the most resistant microorganisms to aPAA among our test organisms, the bacteriophage MS2, and spores of &lt;i&gt;Geobacillus stearothermophilus&lt;/i&gt;. Thus, we are now able to present the first aerosol-based room disinfection method suitable for aircraft that could easily be adopted for other aircraft types and other means of mass transport as well.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Importance: &lt;/strong&gt;Despite numerous outbreaks in recent years, particularly the Ebola virus epidemic and the severe acute respiratory syndrome coronavirus 2 pandemic, there is still no approved method for disinfecting contaminated aircraft. Due to the highly complex nature of aircraft material and technology, it is difficult to balance the antimicrobial efficacy and material compatibility of the biocides used in a way that meets high safety standards. Here, we demonstrate an aerosol-based disinfection protocol using a mixture of peracetic acid and hydrogen peroxide. This protocol passed all material compatibility tests according to relevant aircraft certification specifications established by the European Union Aviation Safety Agency, as well as the manufacturer qualification test programs, and proved to be highly effective against viruses, bacteria, and even bacterial endospores without compromising the airworthiness of the aircraft. By targeting both contaminated surfaces and objects, as well as airborne pathogen reservoirs, it minimizes the spillover risk of zoonotic agents and prevents the mechanical transmission of high-consequence agricultural pathogens into disease-free geographical regions. This makes","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0054226"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13639886/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148866160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum for Pires et al., \"<i>Plasmodium falciparum</i> leucine-rich repeat 5 disruption alters the transcription progression during asexual and sexual stage development\".","authors":"Camilla V Pires, Changqi C Q Wang, Min Zhang, Chiara Micchelli, Prem Prakash, Jenna Oberstaller, Julian C Rayner, Thomas D Otto, John H Adams","doi":"10.1128/msphere.00592-26","DOIUrl":"10.1128/msphere.00592-26","url":null,"abstract":"","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0059226"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621857/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148818973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mSphere of Influence: The age of antifungal resistance.","authors":"Norman van Rhijn","doi":"10.1128/msphere.00009-26","DOIUrl":"10.1128/msphere.00009-26","url":null,"abstract":"<p><p>Norman van Rhijn works in the field of One Health relating to fungal pathogens and antifungal resistance studying the factors that shape the emergence and spread of fungal diseases. In this mSphere of Influence article, they reflect on how the work by M. E. Gorris, K. K. Treseder, C. S. Zender, and J. T. Randerson (GeoHealth 3:308-327, 2019, https://doi.org/10.1029/2019GH0002) made an impact on them by evidencing that environmental change directly impacted the change in habitat of <i>Coccidioides immitis</i>, the fungal pathogen responsible for Valley fever. This work has shaped their approach toward understanding fungal pathogens and environmental change driving antifungal resistance.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0000926"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621836/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148795897","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Serine ubiquitination of the SARS-CoV-2 RdRp by the non-canonical E3 ligase MYCBP2.","authors":"RuthMabel Boytz, Jadon Layne, Cleopatra Skerrit-Genelus, Chengjin Ye, Jack Chun-Chieh Hsu, Luis Martinez-Sobrido, Maudry Laurent-Rolle","doi":"10.1128/msphere.00373-26","DOIUrl":"10.1128/msphere.00373-26","url":null,"abstract":"<p><p>Post-translational modifications, including ubiquitination, have emerged as important regulators of viral infection and the host innate immune response. We screened the proteins of SARS-CoV-2 for ubiquitination and identified the RNA-dependent RNA polymerase (RdRp) NSP12 as being ubiquitinated, which has not previously been reported. Importantly, we confirmed that NSP12 is ubiquitinated during SARS-CoV-2 infection, and the NSP12 proteins of different coronaviruses are differentially associated with ubiquitin chains. SARS-CoV-2 NSP12 was primarily associated with K63-linked polyubiquitin chains, which increased SARS-CoV-2 NSP12 stability relative to OC43 NSP12, which was associated with K48 chains. Additionally, we identify the atypical E3 ubiquitin ligase MYCBP2 as a mediator of serine ubiquitination of SARS-CoV-2 NSP12 on serine-564 and observe that MYCBP2 promotes SARS-CoV-2 replication in cell-based assays. Substitution of this single residue (S564A) in CoV-2 NSP12 prevented recovery of infectious virus in three independent attempts, precluding direct genetic validation of its role in viral replication. To our knowledge, these findings provide the first evidence of serine ubiquitination of a viral protein and suggest a previously unrecognized role for RdRp ubiquitination in coronavirus biology.IMPORTANCEViruses often hijack host post-translational modification systems, including ubiquitination, to enhance the functions of their proteins and evade host innate immune responses. We screened the viral proteins of SARS-CoV-2 and found that NSP2, NSP5, NSP12, NSP15, and NSP16 are ubiquitinated. Focusing on NSP12, the RNA-dependent RNA polymerase (RdRp), we found that it is associated with multiple types of ubiquitin modifications, and RdRp proteins from other coronaviruses are differentially ubiquitinated. We also found that SARS-CoV-2 RdRp undergoes ubiquitination on a serine, rather than a canonical lysine residue, mediated by the atypical E3 ubiquitin ligase, MYCBP2. Consistent with this, MYCBP2 promotes efficient SARS-CoV-2 replication in cell-based assays. To our knowledge, this study provides the first evidence of non-lysine ubiquitination of a viral protein and demonstrates the serine selectivity of MYCBP2 for the first time in a cellular context. These findings raise important questions about the regulatory function of serine NSP12 ubiquitination in coronavirus replication and its potential contribution to viral pathogenicity.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0037326"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621839/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148866098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oxygen underpins the temperature-size relationship of protist plankton.","authors":"Keisuke Inomura, Meng Gao, Moge Du, Curtis Deutsch","doi":"10.1128/msphere.00146-26","DOIUrl":"10.1128/msphere.00146-26","url":null,"abstract":"<p><p>Temperature is known to affect organismal size, yet the underlying mechanism has been debated. Here, we develop a simple growth model of heterotrophic plankton and explain the temperature-size effect based on the demand and supply of oxygen. The model applies the growth rate-temperature relationship based on protein stability, capturing six data sets of a planktonic ciliate <i>Urotricha</i>. The model then computes the oxygen demand, based on the growth rate, which must be balanced by the diffusive uptake of oxygen. Because the smaller cell size favors oxygen uptake per cell volume, the predicted volume-temperature relationship is roughly the inversion of the growth rate-temperature relationship. With this inversive pattern, models are able to capture the data. This model-data consistency indicates that <i>Urotricha</i> may adjust their body size to meet the oxygen demands. Given that similar inversive relationships have been observed in other organisms, we hypothesize that oxygen demand and uptake are the key factors in constraining the volume of heterotrophic planktonic organisms.</p><p><strong>Importance: </strong>The temperature-size relationship is a famous rule in physiological ecology based on common observations, but the mechanism underlying this rule has remained unclear. In this study, we develop a simple growth model of heterotrophic plankton, which explains the temperature-size relationship based on the demand and supply of oxygen. The model predicts temperature change will cause growth rate change; with higher growth rates, the increase of O₂ demand causes a smaller cellular size. The model results also capture six datasets of <i>Urotricha</i> ciliates, suggesting that these organisms may adjust their body size to meet oxygen requirements. Given that similar patterns have been observed in other organisms, we hypothesize that oxygen demand and uptake are key factors constraining cell volume. Our study highlights oxygen as an intermediate factor underpinning temperature and cell volume, providing an explanation for the temperature-size relationship.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0014626"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621864/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148866154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mSphere of Influence: Embracing biological complexity.","authors":"Linda Catherine Horianopoulos","doi":"10.1128/msphere.00218-26","DOIUrl":"10.1128/msphere.00218-26","url":null,"abstract":"<p><p>Linda Horianopoulos is a yeast biologist who uses a combination of comparative genomics, evolutionary analyses, and targeted molecular genetics to research the diversity of metabolism and stress responses across different yeast species. In this mSphere of Influence article, she highlights how a research article about Hsp90's ability to potentiate rapid evolution of drug resistance across different fungal species (https://doi.org/10.1126/science.1118370) influenced her work and the lens through which she appreciates biological complexity.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0021826"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621837/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148764908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Elevated temperature undermines the efficacy of <i>Streptococcus pyogenes</i> dCas9-based CRISPRi systems.","authors":"Lili M Kim, Byoung-Mo Koo, Carol A Gross, Horia Todor","doi":"10.1128/msphere.00379-26","DOIUrl":"10.1128/msphere.00379-26","url":null,"abstract":"<p><p>Targeted gene repression using CRISPRi has become a foundational technique for microbiology. The effector protein in most CRISPRi systems is a catalytically inactivated version of Cas9, an RNA-guided DNA endonuclease from <i>Streptococcus pyogenes</i> (SpCas9). The application of SpCas9-based CRISPRi systems in increasingly ambitious experiments in diverse bacteria raises important questions about the effects of growth temperature on CRISPRi efficacy, particularly because SpCas9 loses cutting efficacy at elevated temperatures. Here, we test the effect of temperature on SpCas9<i>-</i>based CRISPRi knockdown efficacy by performing a series of genome-wide screens using a well-characterized CRISPRi system in the model firmicute <i>Bacillus subtilis</i>. We observe very little repression at high (45°C) growth temperatures and confirm this observation using single cell measurements of GFP fluorescence in CRISPRi strains grown at different temperatures. Our data suggest that <i>Streptococcus pyogenes-</i>based CRISPRi systems are ill-suited for thermophiles and should not be used for elevated-temperature or heat shock experiments in mesophilic bacteria.</p><p><strong>Importance: </strong>CRISPRi is a common technique for inducible gene depletion in bacteria and has been used for many different types of experiments in increasingly diverse bacteria. We identify increased temperatures as an important limitation of the most common CRISPRi effector (<i>Streptococcus pyogenes</i> dCas9). This finding both affects experimental design and informs the kinds of bacterial species amenable to manipulation by <i>S. pyogenes</i>-based CRISPRi systems.</p>","PeriodicalId":19052,"journal":{"name":"mSphere","volume":" ","pages":"e0037926"},"PeriodicalIF":4.0,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13621834/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148795834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}