High-throughput sequencing of Precise CRISPR/Cas9-targeted Recessive Dystrophic Epidermolysis Bullosa immortalised keratinocytes (Ion Torrent)

In this study, the COL7A1 on-target (exon 73) was analysed for precise CRISPR/Cas9 activity.
NGS data was assessed in CRISPR/Cas9-treated immortalised patient keratinocytes
Despite high on-target activity, high homgeneity of DSB repair outcomes was observed at the nuclease target site. This should result in efficient reframing of the gene, with minimal divergence from the wild-type sequence.

在本研究中，对COL7A1靶点（外显子73）进行了精确的CRISPR/Cas9活性分析。对CRISPR/Cas9处理的永生化患者角质形成细胞中的NGS数据进行了评估。尽管靶点活性较高，但在核酸酶靶点位置观察到双链断裂修复结果的同质性较高。这应导致基因的有效重构，与野生型序列的最小差异。