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A thermostable Cas9 with increased lifetime in human plasma
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Journal Article
A thermostable Cas9 with increased lifetime in human plasma
2017
Overview
CRISPR-Cas9 is a powerful technology that has enabled genome editing in a wide range of species. However, the currently developed Cas9 homologs all originate from mesophilic bacteria, making them susceptible to degradation and unsuitable for applications requiring cleavage at elevated temperatures. Here, we show that the Cas9 protein from the thermophilic bacterium
Geobacillus stearothermophilus
(GeoCas9) catalyzes RNA-guided DNA cleavage at elevated temperatures. GeoCas9 is active at temperatures up to 70 °C, compared to 45 °C for
Streptococcus pyogenes
Cas9 (SpyCas9), which expands the temperature range for CRISPR-Cas9 applications. We also found that GeoCas9 is an effective tool for editing mammalian genomes when delivered as a ribonucleoprotein (RNP) complex. Together with an increased lifetime in human plasma, the thermostable GeoCas9 provides the foundation for improved RNP delivery in vivo and expands the temperature range of CRISPR-Cas9.
While current CRISPR-Cas9 tools have revolutionized genome editing, they are not suitable for applications at elevated temperatures. Here, the authors characterize GeoCas9 from
Geobacillus stearothermophilus
, which is active up to 70°C and is stable in human plasma.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ Bacteria
/ Bacterial Proteins - administration & dosage
/ Bacterial Proteins - metabolism
/ Cleavage
/ CRISPR
/ DNA
/ Editing
/ Endonucleases - administration & dosage
/ Enzymes
/ Genomes
/ Geobacillus stearothermophilus - enzymology
/ Homology
/ Humanities and Social Sciences
/ Humans
/ Proteins
/ Ribonucleoproteins - administration & dosage
/ RNA
/ Science
