Homo sapiens Targeted Locus (Loci) - Amplicons from gene editing experiment. Homo sapiens

Cystic fibrosis (CF) bronchial epithelial cells were treated with nanoparticles containing peptide nucleic acid (PNA) and donor DNA designed to coordinate correction of the F508del CF mutation. PCR amplicons at on-target and off-target loci from treated cells were sequenced to assess for gene modification, and for off-target genotoxicity. Samples were analyzed by deep sequencing of alleles in the population using Illumina HiSeq sequencer. This study demonstrated site-specific gene editing in cells of interest with low off-target effects.

本研究将携带肽核酸（peptide nucleic acid, PNA）与用于协同矫正F508del囊性纤维化（Cystic fibrosis, CF）突变的供体DNA的纳米颗粒，作用于囊性纤维化支气管上皮细胞。随后对经上述处理的细胞的靶位点与脱靶位点的PCR扩增子进行测序，以评估基因修饰效果及脱靶遗传毒性。实验采用Illumina HiSeq测序仪对细胞群体中的等位基因开展深度测序以完成样本分析。本研究证实，目标细胞可实现低脱靶效应的位点特异性基因编辑。