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Enhancement of prime editing via xrRNA motif-joined pegRNA

Author

Listed:
  • Guiquan Zhang

    (Model Animal Research Center at Medical School of Nanjing University)

  • Yao Liu

    (Northwest A&F University)

  • Shisheng Huang

    (ShanghaiTech University)

  • Shiyuan Qu

    (ShanghaiTech University)

  • Daolin Cheng

    (Model Animal Research Center at Medical School of Nanjing University)

  • Yuan Yao

    (Zhejiang University)

  • Quanjiang Ji

    (ShanghaiTech University)

  • Xiaolong Wang

    (Northwest A&F University)

  • Xingxu Huang

    (ShanghaiTech University
    Zhejiang Laboratory)

  • Jianghuai Liu

    (Model Animal Research Center at Medical School of Nanjing University)

Abstract

The prime editors (PEs) have shown great promise for precise genome modification. However, their suboptimal efficiencies present a significant technical challenge. Here, by appending a viral exoribonuclease-resistant RNA motif (xrRNA) to the 3′-extended portion of pegRNAs for their increased resistance against degradation, we develop an upgraded PE platform (xrPE) with substantially enhanced editing efficiencies in multiple cell lines. A pan-target average enhancement of up to 3.1-, 4.5- and 2.5-fold in given cell types is observed for base conversions, small deletions, and small insertions, respectively. Additionally, xrPE exhibits comparable edit:indel ratios and similarly minimal off-target editing as the canonical PE3. Of note, parallel comparison of xrPE to the most recently developed epegRNA-based PE system shows their largely equivalent editing performances. Our study establishes a highly adaptable platform of improved PE that shall have broad implications.

Suggested Citation

  • Guiquan Zhang & Yao Liu & Shisheng Huang & Shiyuan Qu & Daolin Cheng & Yuan Yao & Quanjiang Ji & Xiaolong Wang & Xingxu Huang & Jianghuai Liu, 2022. "Enhancement of prime editing via xrRNA motif-joined pegRNA," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-29507-x
    DOI: 10.1038/s41467-022-29507-x
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    References listed on IDEAS

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    Cited by:

    1. Hongyuan Zhang & Jiacheng Ma & Zhaowei Wu & Xiaoyang Chen & Yangyang Qian & Weizhong Chen & Zhipeng Wang & Ya Zhang & Huanhu Zhu & Xingxu Huang & Quanjiang Ji, 2024. "BacPE: a versatile prime-editing platform in bacteria by inhibiting DNA exonucleases," Nature Communications, Nature, vol. 15(1), pages 1-9, December.
    2. Ronghao Chen & Yu Cao & Yajing Liu & Dongdong Zhao & Ju Li & Zhihui Cheng & Changhao Bi & Xueli Zhang, 2023. "Enhancement of a prime editing system via optimal recruitment of the pioneer transcription factor P65," Nature Communications, Nature, vol. 14(1), pages 1-8, December.
    3. Jianli Tao & Daniel E. Bauer & Roberto Chiarle, 2023. "Assessing and advancing the safety of CRISPR-Cas tools: from DNA to RNA editing," Nature Communications, Nature, vol. 14(1), pages 1-16, December.
    4. Jidong Fei & Dongdong Zhao & Caiyi Pang & Ju Li & Siwei Li & Wentao Qiao & Juan Tan & Changhao Bi & Xueli Zhang, 2025. "Mismatch prime editing gRNA increased efficiency and reduced indels," Nature Communications, Nature, vol. 16(1), pages 1-10, December.

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