Fig. 2: CRISPR-mediated gene correction of the ATP7A M1311V mutation and construction of patient-derived isogenic iPS cell lines. | Communications Biology

Fig. 2: CRISPR-mediated gene correction of the ATP7A M1311V mutation and construction of patient-derived isogenic iPS cell lines.

From: CRISPR-mediated gene correction links the ATP7A M1311V mutations with amyotrophic lateral sclerosis pathogenesis in one individual

Fig. 2

a Overview of our personalized medicine approach. b Strategy for correction of the ATP7A M1311V mutation in patient-derived iPS cells using CRISPR-Cas9. The ATP7A point mutation (A to G) is shown in red. The 20-nucleotide single guide RNA (sgRNA) target sequence and the protospacer adjacent motif (PAM) are shown in blue and yellow, respectively. The single-stranded oligodeoxynucleotides (ssODNs), which has 50-bp homology arms, was designed to change the disease-causing mutation to the normal sequence and disrupt the PAM sequence to prevent further Cas9-mediated cleavage after the correction. c Efficiency of homology-directed repair (HDR) for generating the gene-corrected isogenic iPS cell line. d Karyotype analysis of parental (ATP7A-M1311V) and gene-corrected (ATP7A-Cor1 and ATP7A-Cor2) iPS cell lines. e Sanger sequencing to confirm the mutant ATP7A M1311V and normal sequence in iPS cells, NPCs, and MNs.

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