ArticleNature communications2017
Characterizing a thermostable Cas9 for bacterial genome editing and silencing.
Article in Nature communications, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 66 papers.
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Who cites it
66 citing papers in PubMed.
- Elevated temperature undermines the efficacy ofmSphere · 2026Article
- Cross-Strain Transferability of CRISPRi Systems and Design Rules from Laboratory to ClinicalACS synthetic biology · 2026Article
- Article
- Genome editing of phylogenetically distinct bacteria using cross-species retron-mediated recombineering.Nature biotechnology · 2026Article
- Cross-strain transferability of CRISPRi systems and design rules from laboratory to clinicalbioRxiv : the preprint server for biology · 2026Article
- Loop engineering of AtCas9 for effective and broad genome editing.Cell insight · 2025Article
- RepFluo, a Fast Fluorescent In Vitro Assay of Cas9 Activity Exploiting Melting Curve Analysis.ACS omega · 2025Article
- Functional Characterization of Fp2Cas9, a Cold-Adapted Type II-C CRISPR Nuclease fromInternational journal of molecular sciences · 2025Article
- A thermostable Cas9-based genome editing system for thermophilic acetogenic bacteriumApplied and environmental microbiology · 2025Article
- Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR-Cas9 ribonucleoprotein.Nature biotechnology · 2025Article
- The engineering toolbox of Parageobacillus thermoglucosidasius.Applied microbiology and biotechnology · 2025Review
- CRISPR-Cas9-driven genome editing inFrontiers in microbiology · 2025Article
- A single-plasmid-based, easily curable CRISPR/Cas9 system for rapid, iterative genome editing in Pseudomonas putida KT2440.Microbial cell factories · 2024Article
- A highly efficient method for genomic deletion across diverse lengths in thermophilicSynthetic and systems biotechnology · 2024Article
- Harnessing the evolving CRISPR/Cas9 for precision oncology.Journal of translational medicine · 2024Review
- Elucidating the biotechnological potential of the genera Parageobacillus and Saccharococcus through comparative genomic and pan-genome analysis.BMC genomics · 2024Article
- Article
- TemStaPro: protein thermostability prediction using sequence representations from protein language models.Bioinformatics (Oxford, England) · 2024Article
- Expression and Functional Analysis of the Compact ThermophilicInternational journal of molecular sciences · 2023Article
- Characterization of the AcrIIC1 anti‒CRISPR protein for Cas9‒based genome engineering in E. coli.Communications biology · 2023Article
6 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR-Cas9-based genome engineering tools have revolutionized fundamental research and biotechnological exploitation of both eukaryotes and prokaryotes. However, the mesophilic nature of the established Cas9 systems does not allow for applications that require enhanced stability, including engineering at elevated temperatures. Here we identify and characterize ThermoCas9 from the thermophilic bacterium Geobacillus thermodenitrificans T12. We show that in vitro ThermoCas9 is active between 20 and 70 °C, has stringent PAM-preference at lower temperatures, tolerates fewer spacer-protospacer mismatches than SpCas9 and its activity at elevated temperatures depends on the sgRNA-structure. We develop ThermoCas9-based engineering tools for gene deletion and transcriptional silencing at 55 °C in Bacillus smithii and for gene deletion at 37 °C in Pseudomonas putida. Altogether, our findings provide fundamental insights into a thermophilic CRISPR-Cas family member and establish a Cas9-based bacterial genome editing and silencing tool with a broad temperature range.
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