A CRISPR/Cas-mediated selection-free knockin strategy in human embryonic stem cells Journal Article
| Authors: | Zhu, Z.; Verma, N.; González, F.; Shi, Z. D.; Huangfu, D. |
| Article Title: | A CRISPR/Cas-mediated selection-free knockin strategy in human embryonic stem cells |
| Abstract: | The development of new gene-editing tools, in particular the CRISPR/Cas system, has greatly facilitated site-specific mutagenesis in human embryonic stem cells (hESCs), including the introduction or correction of patient-specific mutations for disease modeling. However, integration of a reporter gene into an endogenous locus in hESCs still requires a lengthy and laborious two-step strategy that involves first drug selection to identify correctly targeted clones and then excision of the drug-resistance cassette. Through the use of iCRISPR, an efficient gene-editing platform we recently developed, this study demonstrates a knockin strategy without drug selection for both active and silent genes in hESCs. Lineage-specific hESC reporter lines are useful for real-time monitoring of cell-fate decisions and lineage tracing, as well as enrichment of specific cell populations during hESC differentiation. Thus, this selection-free knockin strategy is expected to greatly facilitate the use of hESCs for developmental studies, disease modeling, and cell-replacement therapy. © 2015 The Authors. |
| Journal Title: | Stem Cell Reports |
| Volume: | 4 |
| Issue: | 6 |
| ISSN: | 2213-6711 |
| Publisher: | Cell Press |
| Date Published: | 2015-06-09 |
| Start Page: | 1103 |
| End Page: | 1111 |
| Language: | English |
| DOI: | 10.1016/j.stemcr.2015.04.016 |
| PROVIDER: | scopus |
| PMCID: | PMC4471821 |
| PUBMED: | 26028531 |
| DOI/URL: | |
| Notes: | Export Date: 2 July 2015 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
MSK Authors
Related MSK Work