Juan-Juan Zhao, Xin-Yu Sun, Sai-Ning Tian, Zong-Ze Zhao, Meng-Di Yin, Mei Zhao, Feng Zhang, Si-Ang Li, Zhi-Xue Yang, Wei Wen, Tao Cheng, An Gong, Jian-Ping Zhang, Xiao-Bing Zhang
BACKGROUND: CRISPR-Cas9 technology has advanced in vivo gene therapy for disorders like hemophilia A, notably through the successful targeted incorporation of the F8 gene into the Alb locus in hepatocytes, effectively curing this disorder in mice. However, thoroughly evaluating the safety and specificity of this therapy is essential. Our study introduces a novel methodology to analyze complex insertion sequences at the on-target edited locus, utilizing barcoded long-range PCR, CRISPR RNP-mediated deletion of unedited alleles, magnetic bead-based long amplicon enrichment, and nanopore sequencing...
February 17, 2024: BMC Genomics