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Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang

Author

Listed:
  • Su Moon

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Jeong Mi Lee

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Jeong Gu Kang

    (Genome Editing Research Center, KRIBB)

  • Nan-Ee Lee

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Dae-In Ha

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Do Yon Kim

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Sun Hee Kim

    (Genome Editing Research Center, KRIBB)

  • Kwangsun Yoo

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Daesik Kim

    (Seoul National University)

  • Jeong-Heon Ko

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

  • Yong-Sam Kim

    (Genome Editing Research Center, KRIBB
    Korea University of Science and Technology (UST))

Abstract

Genome editing has been harnessed through the development of CRISPR system, and the CRISPR from Prevotella and Francisella 1 (Cpf1) system has emerged as a promising alternative to CRISPR-Cas9 for use in various circumstances. Despite the inherent multiple advantages of Cpf1 over Cas9, the adoption of Cpf1 has been unsatisfactory because of target-dependent insufficient indel efficiencies. Here, we report an engineered CRISPR RNA (crRNA) for highly efficient genome editing by Cpf1, which includes a 20-base target-complementary sequence and a uridinylate-rich 3′-overhang. When the crRNA is transcriptionally produced, crRNA with a 20-base target-complementary sequence plus a U4AU4 3′-overhang is the optimal configuration. U-rich crRNA also maximizes the utility of the AsCpf1 mutants and multiplexing genome editing using mRNA as the source of multiple crRNAs. Furthermore, U-rich crRNA enables a highly safe and specific genome editing using Cpf1 in human cells, contributing to the enhancement of a genome-editing toolbox.

Suggested Citation

  • Su Moon & Jeong Mi Lee & Jeong Gu Kang & Nan-Ee Lee & Dae-In Ha & Do Yon Kim & Sun Hee Kim & Kwangsun Yoo & Daesik Kim & Jeong-Heon Ko & Yong-Sam Kim, 2018. "Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang," Nature Communications, Nature, vol. 9(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-06129-w
    DOI: 10.1038/s41467-018-06129-w
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    Cited by:

    1. Xiangfeng Kong & Hainan Zhang & Guoling Li & Zikang Wang & Xuqiang Kong & Lecong Wang & Mingxing Xue & Weihong Zhang & Yao Wang & Jiajia Lin & Jingxing Zhou & Xiaowen Shen & Yinghui Wei & Na Zhong & W, 2023. "Engineered CRISPR-OsCas12f1 and RhCas12f1 with robust activities and expanded target range for genome editing," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    2. Guanhua Xun & Zhixin Zhu & Nilmani Singh & Jingxia Lu & Piyush K. Jain & Huimin Zhao, 2024. "Harnessing noncanonical crRNA for highly efficient genome editing," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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