An efficient gene knock-in strategy using 5′-modified double-stranded DNA donors with short homology arms

Yi Yu, Yijun Guo, Qiqi Tian, Yuanqing Lan, Hugh Yeh, Meng Zhang, Ipek Tasan, Surbhi Jain, Huimin Zhao

Research output: Contribution to journalArticle

Abstract

Here, we report a rapid CRISPR–Cas9-mediated gene knock-in strategy that uses Cas9 ribonucleoprotein and 5′-modified double-stranded DNA donors with 50-base-pair homology arms and achieved unprecedented 65/40% knock-in rates for 0.7/2.5 kilobase inserts, respectively, in human embryonic kidney 293T cells. The identified 5′-end modification led to up to a fivefold increase in gene knock-in rates at various genomic loci in human cancer and stem cells.

Original languageEnglish (US)
JournalNature chemical biology
DOIs
StateAccepted/In press - Dec 23 2019

Fingerprint

Gene Knock-In Techniques
Ribonucleoproteins
Neoplastic Stem Cells
HEK293 Cells
DNA
Base Pairing
Kidney

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

An efficient gene knock-in strategy using 5′-modified double-stranded DNA donors with short homology arms. / Yu, Yi; Guo, Yijun; Tian, Qiqi; Lan, Yuanqing; Yeh, Hugh; Zhang, Meng; Tasan, Ipek; Jain, Surbhi; Zhao, Huimin.

In: Nature chemical biology, 23.12.2019.

Research output: Contribution to journalArticle

Yu, Yi ; Guo, Yijun ; Tian, Qiqi ; Lan, Yuanqing ; Yeh, Hugh ; Zhang, Meng ; Tasan, Ipek ; Jain, Surbhi ; Zhao, Huimin. / An efficient gene knock-in strategy using 5′-modified double-stranded DNA donors with short homology arms. In: Nature chemical biology. 2019.
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