Title |
Rapid characterization of CRISPR-Cas9 protospacer adjacent motif sequence elements
|
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Published in |
Genome Biology, November 2015
|
DOI | 10.1186/s13059-015-0818-7 |
Pubmed ID | |
Authors |
Tautvydas Karvelis, Giedrius Gasiunas, Joshua Young, Greta Bigelyte, Arunas Silanskas, Mark Cigan, Virginijus Siksnys |
Abstract |
To expand the repertoire of Cas9s available for genome targeting, we present a new in vitro method for the simultaneous examination of guide RNA and protospacer adjacent motif (PAM) requirements. The method relies on the in vitro cleavage of plasmid libraries containing a randomized PAM as a function of Cas9-guide RNA complex concentration. Using this method, we accurately reproduce the canonical PAM preferences for Streptococcus pyogenes, Streptococcus thermophilus CRISPR3 (Sth3), and CRISPR1 (Sth1). Additionally, PAM and sgRNA solutions for a novel Cas9 protein from Brevibacillus laterosporus are provided by the assay and are demonstrated to support functional activity in vitro and in plants. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
United Kingdom | 3 | 19% |
France | 2 | 13% |
Taiwan | 1 | 6% |
Spain | 1 | 6% |
Finland | 1 | 6% |
Luxembourg | 1 | 6% |
United States | 1 | 6% |
Australia | 1 | 6% |
Unknown | 5 | 31% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 8 | 50% |
Scientists | 7 | 44% |
Science communicators (journalists, bloggers, editors) | 1 | 6% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 3 | <1% |
Korea, Republic of | 1 | <1% |
Switzerland | 1 | <1% |
China | 1 | <1% |
Belgium | 1 | <1% |
Unknown | 359 | 98% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 68 | 19% |
Researcher | 63 | 17% |
Student > Bachelor | 57 | 16% |
Student > Master | 48 | 13% |
Student > Doctoral Student | 15 | 4% |
Other | 37 | 10% |
Unknown | 78 | 21% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 130 | 36% |
Agricultural and Biological Sciences | 104 | 28% |
Immunology and Microbiology | 11 | 3% |
Chemical Engineering | 7 | 2% |
Engineering | 7 | 2% |
Other | 20 | 5% |
Unknown | 87 | 24% |