Chapter title |
Targeted Transcriptional Repression in Bacteria Using CRISPR Interference (CRISPRi).
|
---|---|
Chapter number | 23 |
Book title |
CRISPR
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2687-9_23 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2686-2, 978-1-4939-2687-9
|
Authors |
Hawkins, John S, Wong, Spencer, Peters, Jason M, Almeida, Ricardo, Qi, Lei S, John S. Hawkins, Spencer Wong, Jason M. Peters, Ricardo Almeida, Lei S. Qi |
Editors |
Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran |
Abstract |
Clustered regularly interspersed short palindromic repeats (CRISPR) interference (CRISPRi) is a powerful technology for sequence-specifically repressing gene expression in bacterial cells. CRISPRi requires only a single protein and a custom-designed guide RNA for specific gene targeting. In Escherichia coli, CRISPRi repression efficiency is high (~300-fold), and there are no observable off-target effects. The method can be scaled up as a general strategy for the repression of many genes simultaneously using multiple designed guide RNAs. Here we provide a protocol for efficient guide RNA design, cloning, and assay of the CRISPRi system in E. coli. In principle, this protocol can be used to construct CRISPRi systems for gene repression in other species of bacteria. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 2 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Scientists | 1 | 50% |
Members of the public | 1 | 50% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 1 | <1% |
China | 1 | <1% |
France | 1 | <1% |
Unknown | 160 | 98% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 33 | 20% |
Researcher | 32 | 20% |
Student > Master | 26 | 16% |
Student > Bachelor | 17 | 10% |
Professor > Associate Professor | 9 | 6% |
Other | 13 | 8% |
Unknown | 33 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 57 | 35% |
Agricultural and Biological Sciences | 35 | 21% |
Immunology and Microbiology | 10 | 6% |
Medicine and Dentistry | 4 | 2% |
Chemistry | 4 | 2% |
Other | 14 | 9% |
Unknown | 39 | 24% |