CRISPR-Cas9 mediated genome editing of the parasitic nematode Nippostrongylus brasiliensis

Parasitic nematodes are a major burden for human populations and agriculture, particularly in the developing world. Advances in treatment require better understanding of gene function, but so far approaches to perform functional genomics in parasitic nematodes have been largely unsuccessful. We show CRISPR-Cas9 mediated gene editing in the parasitic nematode N. brasiliensis. We edited the endogenous DNAseII locus to insert a non-functional copy and showed that this reduced expression of DNAseII. High throughput sequencing, presented here of the edited locus in N. brasiliensis genomic DNA confirmed that the endogenous locus was replaced by the non-functional copy with almost 100% accuracy.

寄生线虫（Parasitic nematodes）对人类社会与农业生产造成了沉重负担，在发展中地区尤为突出。治疗手段的研发升级有赖于对其基因功能的深入解析，但迄今针对寄生线虫开展功能基因组学研究的方法大多未能取得有效进展。本研究证实，CRISPR-Cas9系统（CRISPR-Cas9）介导的基因编辑技术可应用于寄生线虫巴西日圆线虫（Nippostrongylus brasiliensis，简称N. brasiliensis）。我们通过编辑其内源脱氧核糖核酸酶II（DNAse II）基因座，插入一段无功能拷贝，并证实该操作可降低脱氧核糖核酸酶II的表达水平。本研究中针对巴西日圆线虫基因组DNA内编辑位点的高通量测序（High-throughput sequencing）结果证实，内源基因座已被无功能拷贝精准替换，替换准确率接近100%。