利用CRISPR/Cas9技术对人多能干细胞进行高效基因组编辑

doi:10.16288/j.yczz.15-240

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Hereditas(Beijing) ›› 2015, Vol. 37 ›› Issue (11): 1167-1173.doi: 10.16288/j.yczz.15-240

Efficient genome editing in human pluripotent stem cells through CRISPR/Cas9

Gaigai Liu,Shuang Li,Yuda Wei,Yongxian Zhang,Qiurong Ding

Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Shanghai 200031, China

Received:2015-05-29 Revised:2015-06-24 Online:2015-11-20 Published:2015-06-29

Abstract

Abstract: The RNA-guided CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease has offered a new platform for genome editing with high efficiency. Here, we report the use of CRISPR/Cas9 technology to target a specific genomic region in human pluripotent stem cells. We show that CRISPR/Cas9 can be used to disrupt a gene by introducing frameshift mutations to gene coding region; to knock in specific sequences (e.g. FLAG tag DNA sequence) to targeted genomic locus via homology directed repair; to induce large genomic deletion through dual-guide multiplex. Our results demonstrate the versatile application of CRISPR/Cas9 in stem cell genome editing, which can be widely utilized for functional studies of genes or genome loci in human pluripotent stem cells.

Key words: CRISPR/Cas9 genome editing, human pluripotent stem cells, gene disruption, targeted DNA insertion, largegenomic deletion

Gaigai Liu,Shuang Li,Yuda Wei,Yongxian Zhang,Qiurong Ding. Efficient genome editing in human pluripotent stem cells through CRISPR/Cas9[J]. Hereditas(Beijing), 2015, 37(11): 1167-1173.

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Efficient genome editing in human pluripotent stem cells through CRISPR/Cas9

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