Homo sapiens Targeted Locus (Loci)

The CRISPR/Cas9 genome editing platform is a promising technology to correct the genetic basis of hereditary diseases. The high versatility, efficiency, and multiplexing capabilities of the CRISPR/Cas9 system enable a variety of otherwise challenging gene correction strategies. Here we used the CRISPR/Cas9 system to restore the expression of the dystrophin gene in cells carrying dystrophin mutations that cause Duchenne muscular dystrophy (DMD). We designed single or multiplexed single guide RNAss (sgRNAs) to restore the dystrophin reading frame by targeting the mutational hotspot at exons 45-55 and introducing shifts within exons or deleting one or more exons. Deep sequencing of on-target and off-target sites was used to quantify gene editing activity with high sensitivity.