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CtIP fusion to Cas9 enhances transgene integration by homology-dependent repair.

Abstract：

:In genome editing with CRISPR-Cas9, transgene integration often remains challenging. Here, we present an approach for increasing the efficiency of transgene integration by homology-dependent repair (HDR). CtIP, a key protein in early steps of homologous recombination, is fused to Cas9 and stimulates transgene integration by HDR at the human AAVS1 safe harbor locus. A minimal N-terminal fragment of CtIP, designated HE for HDR enhancer, is sufficient to stimulate HDR and this depends on CDK phosphorylation sites and the multimerization domain essential for CtIP activity in homologous recombination. HDR stimulation by Cas9-HE, however, depends on the guide RNA used, a limitation that may be overcome by testing multiple guides to the locus of interest. The Cas9-HE fusion is simple to use and allows obtaining twofold or more efficient transgene integration than that with Cas9 in several experimental systems, including human cell lines, iPS cells, and rat zygotes.

journal_name

Nat Commun

journal_title

Nature communications

authors

Charpentier M,Khedher AHY,Menoret S,Brion A,Lamribet K,Dardillac E,Boix C,Perrouault L,Tesson L,Geny S,De Cian A,Itier JM,Anegon I,Lopez B,Giovannangeli C,Concordet JP

doi

10.1038/s41467-018-03475-7

subject

Has Abstract

pub_date

2018-03-19 00:00:00

pages

1133

issue

1

issn

2041-1723

pii

10.1038/s41467-018-03475-7

journal_volume

9

pub_type

杂志文章

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