| Chapter title |
Genome Engineering Using CRISPR-Cas9 System
|
|---|---|
| Chapter number | 10 |
| Book title |
Chromosomal Mutagenesis
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-1862-1_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-1861-4, 978-1-4939-1862-1
|
| Authors |
Le Cong, Feng Zhang |
| Abstract |
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system is an adaptive immune system that exists in a variety of microbes. It could be engineered to function in eukaryotic cells as a fast, low-cost, efficient, and scalable tool for manipulating genomic sequences. In this chapter, detailed protocols are described for harnessing the CRISPR-Cas9 system from Streptococcus pyogenes to enable RNA-guided genome engineering applications in mammalian cells. We present all relevant methods including the initial site selection, molecular cloning, delivery of guide RNAs (gRNAs) and Cas9 into mammalian cells, verification of target cleavage, and assays for detecting genomic modification including indels and homologous recombination. These tools provide researchers with new instruments that accelerate both forward and reverse genetics efforts. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 8 | <1% |
| United Kingdom | 3 | <1% |
| Canada | 3 | <1% |
| Italy | 3 | <1% |
| France | 2 | <1% |
| Japan | 2 | <1% |
| Germany | 2 | <1% |
| Belgium | 2 | <1% |
| Bulgaria | 1 | <1% |
| Other | 11 | <1% |
| Unknown | 1711 | 98% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 384 | 22% |
| Student > Bachelor | 321 | 18% |
| Student > Master | 268 | 15% |
| Researcher | 236 | 14% |
| Student > Doctoral Student | 79 | 5% |
| Other | 194 | 11% |
| Unknown | 266 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 621 | 36% |
| Agricultural and Biological Sciences | 465 | 27% |
| Medicine and Dentistry | 91 | 5% |
| Neuroscience | 58 | 3% |
| Immunology and Microbiology | 45 | 3% |
| Other | 166 | 9% |
| Unknown | 302 | 17% |