In vivo genome editing by NanoMEDIC CRISPR-Cas9/sgRNA delivery into a Luc-reporter mosue model.

We first generated a transgenic luciferase reporter mouse model to monitor human Dystrophin gene exon 45 skipping activity. Two CRISPR-sgRNAs and Cas9 protein were delivered by NanoMEDIC technology into gastrocnemius muscle of the reporter mice, and genomic DNAs were extracted 189 days post-injection. The target region was PCR amplified and subjected to Illumina MiSeq amplicon sequencing (300 bp ×2). Each sample (PBS injection: 3 mice, NanoMEDIC injection: 3 mice) was spitted by unique 4 bp index, and pair-end reads were merged by FLASH2 software.