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TALEN outperforms Cas9 in editing heterochromatin target sites

Author

Listed:
  • Surbhi Jain

    (University of Illinois at Urbana−Champaign)

  • Saurabh Shukla

    (University of Illinois at Urbana−Champaign
    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
    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
    University of Illinois at Urbana-Champaign
    University of Illinois at Urbana−Champaign)

  • Paul R. Selvin

    (University of Illinois at Urbana−Champaign
    University of Illinois at Urbana−Champaign
    University of Illinois at Urbana−Champaign)

  • Huimin Zhao

    (University of Illinois at Urbana−Champaign
    University of Illinois at Urbana−Champaign
    University of Illinois at Urbana−Champaign
    University of Illinois at Urbana−Champaign)

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.

Suggested Citation

  • 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 & Hui, 2021. "TALEN outperforms Cas9 in editing heterochromatin target sites," Nature Communications, Nature, vol. 12(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-020-20672-5
    DOI: 10.1038/s41467-020-20672-5
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