Fig. 5: HDR-mediated knock-in via the CRISPR-VIM method with SpCas9/sgRNA packaged in VLPs. | Nature Communications

Fig. 5: HDR-mediated knock-in via the CRISPR-VIM method with SpCas9/sgRNA packaged in VLPs.

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

Fig. 5: HDR-mediated knock-in via the CRISPR-VIM method with SpCas9/sgRNA packaged in VLPs.

a Schematic of the CRISPR-VIM-based HDR strategy with AAV-based donors in mouse embryos. Created in BioRender. Kim, K. (2025) https://BioRender.com/a99d722. b Strategy for generating a knock-in mouse model where exon 5 and flanking intronic sequences (a total of 728 bp) of the mouse Kcnq4 gene are replaced with human KCNQ4 (hKCNQ4) pathogenic mutation and sequences. c PCR-based genotyping of Kcnq4 knock-in mouse embryos, confirming the introduction of human sequences via the CRISPR-VIM-based HDR approach. d Genotyping analysis of F0 mice demonstrating the successful knock-in of human sequences at the Kcnq4 locus. e Sanger sequencing of F0 mice validating the precise knock-in of human sequences into the Kcnq4 locus. f Germline transmission analysis of humanized KCNQ4 mouse genotypes, shown via gel electrophoresis (left) and Sanger sequencing (right). WT indicates wild-type, and KI indicates knock-in. All experiments were repeated three times independently. Source data are provided as a Source Data file.

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