PT  - JOURNAL ARTICLE
AU  - Keun-Tae Kim
AU  - Ju-Chan Park
AU  - Haeseung Lee
AU  - Hyeon-Ki Jang
AU  - Yan Jin
AU  - Wankyu Kim
AU  - Jeongmi Lee
AU  - Hyongbum Henry Kim
AU  - Sang-Su Bae
AU  - Hyuk-Jin Cha
TI  - Scarless Enriched selection of Genome edited Human Pluripotent Stem Cells Using Induced Drug Resistance
AID  - 10.1101/522383
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 522383
4099  - http://biorxiv.org/content/early/2019/01/18/522383.short
4100  - http://biorxiv.org/content/early/2019/01/18/522383.full
AB  - An efficient gene editing technique for use in human pluripotent stem cells (hPSCs) would have great potential value in regenerative medicine, as well as in drug discovery based on isogenic human disease models. However, the extremely low efficiency of gene editing in hPSCs is a major technical hurdle that remains to be resolved. Previously, we demonstrated that YM155, a survivin inhibitor developed as an anti-cancer drug, induces highly selective cell death in undifferentiated hPSCs. In this study, we demonstrated that the high cytotoxicity of YM155 in hPSCs, which is mediated by selective cellular uptake of the drug, is due to high expression of SLC35F2 in these cells. Consistent with this, knockout of SLC35F2 with CRISPR-Cas9 or depletion with siRNAs made hPSCs highly resistant to YM155. Simultaneous gene editing of a gene of interest and transient knockdown of SLC35F2 following YM155 treatment enabled genome-edited hPSCs to survive because YM155 resistance was temporarily induced, thereby achieving enriched selection of genome-edited clonal populations. This precise and efficient genome editing approach took as little as 3 weeks without cell sorting or introduction of additional genes.