Chapter title |
Establishment of the CRISPR/Cas9 System for Targeted Gene Disruption and Gene Tagging
|
---|---|
Chapter number | 11 |
Book title |
Functional Genomics
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7231-9_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7230-2, 978-1-4939-7231-9
|
Authors |
Eric Ehrke-Schulz, Maren Schiwon, Claudia Hagedorn, Anja Ehrhardt |
Abstract |
CRISPR/Cas9 RNA-guided nucleases refashioned in vivo gene editing approaches for specific gene disruption, gene correction, or gene addition. Moreover, chimeric Cas9 proteins can be applied to direct fused cis-acting effector protein domains, enzymes, or fluorescent markers to DNA to target sequences to regulate gene expression, to introduce epigenetic changes, or to fluorescently label DNA sequences of interest. Here we show how to design guide RNAs for specific DNA targeting. We provide a protocol to customize the CRISPR/Cas9 machinery encoded on commercially available plasmids and present how to test the targeting efficiency of Cas9 with a target-specific gRNA by testing mutation induction efficiency. To exemplify related applications we provide a guideline of how to apply the CRISPR/Cas9 technology for gene labeling. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Unknown | 2 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 11 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 3 | 27% |
Professor > Associate Professor | 1 | 9% |
Student > Bachelor | 1 | 9% |
Researcher | 1 | 9% |
Unknown | 5 | 45% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 18% |
Agricultural and Biological Sciences | 2 | 18% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 9% |
Engineering | 1 | 9% |
Unknown | 5 | 45% |