TALEN Outperforms Cas9 in Editing Heterochromatin Target Sites

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Jan 28

PMID 33504770

Citations 41

Authors

Surbhi Jain

Saurabh Shukla

Che Yang

Meng Zhang

Zia Fatma

Manasi Lingamaneni

Shireen Abesteh

Stephan Thomas Lane

Xiong Xiong

Yuchuan Wang

Charles M Schroeder

Paul R Selvin

Huimin Zhao

Affiliations

Soon will be listed here.

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.

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