CRISPR/Cas9 - Mediated Precise Targeted Integration In Vivo Using a Double Cut Donor with Short Homology Arms.

Yao, Xuan; Wang, Xing; Liu, Junlai; Hu, Xinde; Shi, Linyu; Shen, Xiaowen; Ying, Wenqin; Sun, Xinyao et al. · EBioMedicine · 2017

basic_science · Level V

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Abstract

Precisely targeted genome editing is highly desired for clinical applications. However, the widely used homology-directed repair (HDR)-based genome editing strategies remain inefficient for certain in vivo applications. We here demonstrate a microhomology-mediated end-joining (MMEJ)-based strategy for precisely targeted gene integration in transfected neurons and hepatocytes in vivo with efficiencies up to 20%, much higher (up to 10 fold) than HDR-based strategy in adult mouse tissues. As a proof of concept of its therapeutic potential, we demonstrate the efficacy of MMEJ-based strategy in correction of Fah mutation and rescue of Fah<sup>-/-</sup> liver failure mice, offering an efficient approach for precisely targeted gene therapies.

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