CRISPR/Cas-mediated knockin in human pluripotent stem cells Journal Article
| Authors: | Verma, N.; Zhu, Z.; Huangfu, D. |
| Article Title: | CRISPR/Cas-mediated knockin in human pluripotent stem cells |
| 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 effi ciency that necessitates a lengthy process of selection followed by the removal of the selection cassette. Here we describe a procedure to generate fl uorescent reporter and epitope tagged hPSCs in an effi cient 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. © Springer Science+Business Media New York 2017. |
| Keywords: | homologous recombination; gene targeting; fluorescent reporter; crispr/cas; knockin; human pluripotent stem cells (hpscs); epitope tag |
| Journal Title: | Methods in Molecular Biology |
| Volume: | 1513 |
| ISSN: | 1064-3745 |
| Publisher: | Humana Press Inc |
| Date Published: | 2017-01-01 |
| Start Page: | 119 |
| End Page: | 140 |
| Language: | English |
| DOI: | 10.1007/978-1-4939-6539-7_9 |
| PROVIDER: | scopus |
| PUBMED: | 27807834 |
| PMCID: | PMC5554709 |
| DOI/URL: | |
| Notes: | Book Chapter 9 in "Cancer Gene Networks" (ISBN: 978-1-4939-6539-7) -- Export Date: 6 December 2016 -- Source: Scopus |
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