Title |
A thermostable Cas9 with increased lifetime in human plasma
|
---|---|
Published in |
Nature Communications, November 2017
|
DOI | 10.1038/s41467-017-01408-4 |
Pubmed ID | |
Authors |
Lucas B. Harrington, David Paez-Espino, Brett T. Staahl, Janice S. Chen, Enbo Ma, Nikos C. Kyrpides, Jennifer A. Doudna |
Abstract |
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. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 24 | 24% |
Japan | 13 | 13% |
United Kingdom | 12 | 12% |
France | 4 | 4% |
Netherlands | 4 | 4% |
Germany | 2 | 2% |
India | 2 | 2% |
Switzerland | 2 | 2% |
Finland | 1 | 1% |
Other | 4 | 4% |
Unknown | 31 | 31% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 69 | 70% |
Scientists | 26 | 26% |
Science communicators (journalists, bloggers, editors) | 4 | 4% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Switzerland | 1 | <1% |
Unknown | 274 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 55 | 20% |
Researcher | 48 | 17% |
Student > Bachelor | 31 | 11% |
Student > Master | 23 | 8% |
Other | 16 | 6% |
Other | 34 | 12% |
Unknown | 68 | 25% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 96 | 35% |
Agricultural and Biological Sciences | 54 | 20% |
Chemistry | 12 | 4% |
Medicine and Dentistry | 9 | 3% |
Immunology and Microbiology | 7 | 3% |
Other | 25 | 9% |
Unknown | 72 | 26% |