| Chapter title |
CRISPR-Cas9-Mediated Gene Editing in Mouse Spermatogonial Stem Cells
|
|---|---|
| Chapter number | 20 |
| Book title |
RNAi and Small Regulatory RNAs in Stem Cells
|
| Published in |
Methods in molecular biology, July 2017
|
| DOI | 10.1007/978-1-4939-7108-4_20 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7106-0, 978-1-4939-7108-4
|
| Authors |
Yinghua Wang, Yifu Ding, Jinsong Li, Wang, Yinghua, Ding, Yifu, Li, Jinsong |
| Editors |
Baohong Zhang |
| Abstract |
Precise genome editing is a powerful tool for analysis of gene function. However, in spermatogonial stem cells (SSCs), this still remains a big challenge mainly due to low efficiency and complexity of currently available gene editing techniques. The CRISPR-Cas9 system from bacteria has been applied to modifying genome in different species at a very high efficiency and specificity. Here we describe CRISPR-Cas9-mediated gene editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in SSCs. This protocol provides guidelines for derivation of SSCs, nucleofection of SSCs with the CRISPR-Cas9 system, transplantation of the gene-modified SSCs into the recipient testes, and production of mice using transplanted SSC-derived round spermatids. |
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 % |
|---|---|---|
| Unknown | 21 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 3 | 14% |
| Student > Doctoral Student | 2 | 10% |
| Student > Bachelor | 2 | 10% |
| Other | 2 | 10% |
| Student > Ph. D. Student | 2 | 10% |
| Other | 3 | 14% |
| Unknown | 7 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 6 | 29% |
| Agricultural and Biological Sciences | 5 | 24% |
| Immunology and Microbiology | 1 | 5% |
| Engineering | 1 | 5% |
| Unknown | 8 | 38% |