TALEN outperforms Cas9 in editing heterochromatin target sites

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 and Huimin Zhao ()
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Surbhi Jain: University of Illinois at Urbana−Champaign
Saurabh Shukla: University of Illinois at Urbana−Champaign
Che Yang: University of Illinois at Urbana−Champaign
Meng Zhang: University of Illinois at Urbana−Champaign
Zia Fatma: University of Illinois at Urbana−Champaign
Manasi Lingamaneni: University of Illinois at Urbana−Champaign
Shireen Abesteh: University of Illinois at Urbana−Champaign
Stephan Thomas Lane: University of Illinois at Urbana−Champaign
Xiong Xiong: University of Illinois at Urbana−Champaign
Yuchuan Wang: Carnegie Mellon University
Charles M. Schroeder: University of Illinois at Urbana−Champaign
Paul R. Selvin: University of Illinois at Urbana−Champaign
Huimin Zhao: University of Illinois at Urbana−Champaign

Nature Communications, 2021, vol. 12, issue 1, 1-10

Abstract: 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.

Date: 2021
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-020-20672-5

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DOI: 10.1038/s41467-020-20672-5

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