| Chapter title |
CRISPR/Cas-Mediated Knockin in Human Pluripotent Stem Cells.
|
|---|---|
| Chapter number | 9 |
| Book title |
Cancer Gene Networks
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6539-7_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6537-3, 978-1-4939-6539-7
|
| Authors |
Nipun Verma, Zengrong Zhu, Danwei Huangfu, Verma, Nipun, Zhu, Zengrong, Huangfu, Danwei |
| Editors |
Usha Kasid, Robert Clarke |
| Abstract |
Fluorescent reporter and epitope-tagged human pluripotent stem cells (hPSCs) greatly facilitate studies on the pluripotency and differentiation characteristics of these cells. Unfortunately traditional procedures to generate such lines are hampered by a low targeting efficiency that necessitates a lengthy process of selection followed by the removal of the selection cassette. Here we describe a procedure to generate fluorescent reporter and epitope tagged hPSCs in an efficient one-step process using the CRISPR/Cas technology. Although the method described uses our recently developed iCRISPR platform, the protocols can be adapted for general use with CRISPR/Cas or other engineered nucleases. The transfection procedures described could also be used for additional applications, such as overexpression or lineage tracing studies. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 49 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Bachelor | 10 | 20% |
| Researcher | 8 | 16% |
| Student > Ph. D. Student | 6 | 12% |
| Student > Master | 4 | 8% |
| Student > Doctoral Student | 3 | 6% |
| Other | 6 | 12% |
| Unknown | 12 | 24% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 15 | 31% |
| Agricultural and Biological Sciences | 7 | 14% |
| Engineering | 4 | 8% |
| Neuroscience | 3 | 6% |
| Immunology and Microbiology | 2 | 4% |
| Other | 7 | 14% |
| Unknown | 11 | 22% |