Journal: Molecular Therapy. Nucleic Acids

Article Title: Exploring exon excision as a therapeutic intervention strategy for the future treatment of ADGRV1- associated retinitis pigmentosa

doi: 10.1016/j.omtn.2025.102702

Figure Lengend Snippet: Assessment of CRISPR-Cas9 mediated editing efficiency and successful genomic excision of ADGRV1 exons 40–42 in HEK293T cells (A) Assessment of genome-editing efficiency using TIDE analysis following HEK293T transfection with PX459 CRISPR-Cas9 expression plasmids containing sgRNAs targeting either ADGRV1 intron 39 or intron 42 ( n = 2 biological replicates per target). (B) The size of the genomic PCR fragments generated using primers in introns 39 and 42 confirmed the successful excision of ADGRV1 exons 40–42 from HEK293T cell DNA following co-transfection with both plasmids (unedited amplicon size: 3,997 bp; amplicon size after genomic exon excision: 1,950 bp). GAPDH amplification is shown as a loading control (amplicon size: 110 bp). PCR (−), negative PCR control. (C) Sanger sequencing of amplicons confirmed the successful removal of target exons.

Article Snippet: To generate the PX459 plasmids containing the sgRNA sequences, the PX459 plasmid (Addgene #101715) was first linearized with BbsI.

Techniques: CRISPR, Transfection, Expressing, Generated, Cotransfection, Amplification, Control, Sequencing