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Targeted activation of diverse CRISPR-Cas systems for mammalian genome editing via proximal CRISPR targeting.

Abstract：

:Bacterial CRISPR-Cas systems comprise diverse effector endonucleases with different targeting ranges, specificities and enzymatic properties, but many of them are inactive in mammalian cells and are thus precluded from genome-editing applications. Here we show that the type II-B FnCas9 from Francisella novicida possesses novel properties, but its nuclease function is frequently inhibited at many genomic loci in living human cells. Moreover, we develop a proximal CRISPR (termed proxy-CRISPR) targeting method that restores FnCas9 nuclease activity in a target-specific manner. We further demonstrate that this proxy-CRISPR strategy is applicable to diverse CRISPR-Cas systems, including type II-C Cas9 and type V Cpf1 systems, and can facilitate precise gene editing even between identical genomic sites within the same genome. Our findings provide a novel strategy to enable use of diverse otherwise inactive CRISPR-Cas systems for genome-editing applications and a potential path to modulate the impact of chromatin microenvironments on genome modification.

journal_name

Nat Commun

journal_title

Nature communications

authors

Chen F,Ding X,Feng Y,Seebeck T,Jiang Y,Davis GD

doi

10.1038/ncomms14958

subject

Has Abstract

pub_date

2017-04-07 00:00:00

pages

14958

issn

2041-1723

pii

ncomms14958

journal_volume

8

pub_type

杂志文章

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