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An innovative approach using CRISPR-ribonucleoprotein packaged in virus-like particles to generate genetically engineered mouse models

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  • Tae Yeong Jeong

    (Korea University College of Medicine
    Korea University College of Medicine
    Seoul National University)

  • Da Eun Yoon

    (Korea University College of Medicine
    Korea University College of Medicine
    Seoul National University
    Max-Planck Institute of Biochemistry)

  • Sol Pin Kim

    (Seoul National University
    Seoul National University)

  • Jiyun Yang

    (Korea University College of Medicine
    Korea University College of Medicine
    Seoul National University)

  • Soo-Yeon Lim

    (Seoul National University)

  • Sungjin Ok

    (Korea University College of Medicine)

  • Sungjin Ju

    (Korea University College of Medicine
    Korea University College of Medicine
    Seoul National University)

  • Jeongeun Park

    (Korea University College of Medicine
    Seoul National University
    Seoul National University)

  • Su Bin Lee

    (Seoul National University)

  • Soo-Ji Park

    (Korea University College of Medicine
    Korea University College of Medicine
    Seoul National University)

  • Sanghun Kim

    (Korea University College of Medicine
    Korea University College of Medicine)

  • Hyunji Lee

    (Korea University College of Medicine
    Korea University College of Medicine)

  • Daekee Lee

    (Ewha Womans University)

  • Soo Kyung Kang

    (Seoul National University
    Seoul National University)

  • Seung Eun Lee

    (Korea Institute of Science and Technology (KIST))

  • Hyeon Soo Kim

    (Korea University College of Medicine)

  • Je Kyung Seong

    (Seoul National University
    Seoul National University
    Seoul National University
    Seoul National University Cancer Research Institute)

  • Kyoungmi Kim

    (Korea University College of Medicine
    Korea University College of Medicine
    Seoul National University
    Seoul National University)

Abstract

Genetically engineered mouse models (GEMMs) are crucial for investigating disease mechanisms, developing therapeutic strategies, and advancing fundamental biological research. While CRISPR gene editing has greatly facilitated the creation of these models, existing techniques still present technical challenges and efficiency limitations. Here, we establish a CRISPR-VLP-induced targeted mutagenesis (CRISPR-VIM) strategy, enabling precise genome editing by co-culturing zygotes with virus-like particle (VLP)-delivered gene editing ribonucleoproteins (RNPs) without requiring physical manipulation or causing cellular damage. We generate Plin1- and Tyr-knockout mice through VLP-based SpCas9 or adenine base editor (ABE)/sgRNA RNPs and characterize their phenotype and germline transmission. Additionally, we demonstrate cytosine base editor (CBE)/sgRNA-based C-to-T substitution or SpCas9/sgRNA-based knock-in using VLPs. This method further simplifies and accelerates GEMM generation without specialized techniques or equipment. Consequently, the CRISPR-VIM method can facilitate mouse modeling and be applied in various research fields.

Suggested Citation

  • Tae Yeong Jeong & Da Eun Yoon & Sol Pin Kim & Jiyun Yang & Soo-Yeon Lim & Sungjin Ok & Sungjin Ju & Jeongeun Park & Su Bin Lee & Soo-Ji Park & Sanghun Kim & Hyunji Lee & Daekee Lee & Soo Kyung Kang & , 2025. "An innovative approach using CRISPR-ribonucleoprotein packaged in virus-like particles to generate genetically engineered mouse models," Nature Communications, Nature, vol. 16(1), pages 1-13, December.
  • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-58364-7
    DOI: 10.1038/s41467-025-58364-7
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