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

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, Seung Eun Lee, Hyeon Soo Kim, Je Kyung Seong () and Kyoungmi Kim ()
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Tae Yeong Jeong: Korea University College of Medicine
Da Eun Yoon: Korea University College of Medicine
Sol Pin Kim: Seoul National University
Jiyun Yang: Korea University College of Medicine
Soo-Yeon Lim: Seoul National University
Sungjin Ok: Korea University College of Medicine
Sungjin Ju: Korea University College of Medicine
Jeongeun Park: Korea University College of Medicine
Su Bin Lee: Seoul National University
Soo-Ji Park: Korea University College of Medicine
Sanghun Kim: Korea University College of Medicine
Hyunji Lee: Korea University College of Medicine
Daekee Lee: Ewha Womans University
Soo Kyung Kang: 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
Kyoungmi Kim: Korea University College of Medicine

Nature Communications, 2025, vol. 16, issue 1, 1-13

Abstract: 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.

Date: 2025
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DOI: 10.1038/s41467-025-58364-7

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