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TALEN outperforms Cas9 in editing heterochromatin target sites.

Abstract：

:Genome editing critically relies on selective recognition of target sites. However, despite recent progress, the underlying search mechanism of genome-editing proteins is not fully understood in the context of cellular chromatin environments. Here, we use single-molecule imaging in live cells to directly study the behavior of CRISPR/Cas9 and TALEN. Our single-molecule imaging of genome-editing proteins reveals that Cas9 is less efficient in heterochromatin than TALEN because Cas9 becomes encumbered by local searches on non-specific sites in these regions. We find up to a fivefold increase in editing efficiency for TALEN compared to Cas9 in heterochromatin regions. Overall, our results show that Cas9 and TALEN use a combination of 3-D and local searches to identify target sites, and the nanoscopic granularity of local search determines the editing outcomes of the genome-editing proteins. Taken together, our results suggest that TALEN is a more efficient gene-editing tool than Cas9 for applications in heterochromatin.

journal_name

Nat Commun

journal_title

Nature communications

authors

Jain S,Shukla S,Yang C,Zhang M,Fatma Z,Lingamaneni M,Abesteh S,Lane ST,Xiong X,Wang Y,Schroeder CM,Selvin PR,Zhao H

doi

10.1038/s41467-020-20672-5

subject

Has Abstract

pub_date

2021-01-27 00:00:00

pages

606

issue

1

issn

2041-1723

pii

10.1038/s41467-020-20672-5

journal_volume

12

pub_type

杂志文章

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