Efficient scarless genome editing in human pluripotent stem cells. (December 2018)

• Record Type: Journal Article
• Authors: Ikeda, Kazuya; Uchida, Nobuko; Nishimura, Toshinobu; White, Joseph; Martin, Renata; Nakauchi, Hiromitsu; Sebastiano, Vittorio; Weinberg, Kenneth; Porteus, Matthew
• Is Part Of: Nature methods. Volume 15:Number 12(2018)
• Page Start: 1045
• Page End: 1047
• Publication Date: 2018-12
• Subjects: Life sciences -- Methodology -- Periodicals; Life sciences -- Research -- Periodicals; Biology -- Methodology -- Periodicals; Biology -- Research -- Periodicals; 570.72
• DOI: 10.1038/s41592-018-0212-y
• Languages: English
• ISSNs: 1548-7091

Abstract

Abstract Scarless genome editing in human pluripotent stem cells (hPSCs) represents a goal for both precise research applications and clinical translation of hPSC-derived therapies. Here we established a versatile and efficient method that combines CRISPR–Cas9-mediated homologous recombination with positive–negative selection of edited clones to generate scarless genetic changes in hPSCs. The combination of positive and negative selection strategies, paired with the use of shRNAs to avoid random integration, allows efficient and scarless CRISPR-based homologous recombination.