Chapter title |
Analysis of CRISPR Pre-crRNA Cleavage
|
---|---|
Chapter number | 3 |
Book title |
CRISPR
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2687-9_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2686-2, 978-1-4939-2687-9
|
Authors |
Erin L. Garside, Andrew M. MacMillan |
Editors |
Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran |
Abstract |
We have examined the processing of precursor-clustered regularly interspaced short palindromic repeat (CRISPR) RNAs (pre-crRNAs) of the Type I CRISPR-Cas system by incubation of radiolabeled model RNAs with recombinant CRISPR-associated (Cas) endoribonucleases, followed by denaturing polyacrylamide gel electrophoresis (PAGE) of the products. Determination of cleavage position is based on comparison with RNase T1 digestion and base hydrolysis products. The mechanism of cleavage is investigated by chemical and enzymatic characterization of the reaction products as well as by the demonstration that a specific 2'-deoxy substitution 5' to the scissile phosphate blocks endonucleolytic cleavage. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 12 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 2 | 17% |
Student > Master | 2 | 17% |
Researcher | 2 | 17% |
Professor > Associate Professor | 1 | 8% |
Unknown | 5 | 42% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 4 | 33% |
Agricultural and Biological Sciences | 3 | 25% |
Unknown | 5 | 42% |