Department of Microbiology, Immunology & Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, DC 20037, USA.
Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, Giessen, Germany.
Cell Rep Methods. 2023 Jul 24;3(7):100535. doi: 10.1016/j.crmeth.2023.100535.
The identification and characterization of genomic safe harbor sites (GSHs) can facilitate consistent transgene activity with minimal disruption to the host cell genome. We combined computational genome annotation and chromatin structure analysis to predict the location of four GSHs in the human blood fluke, , a major infectious pathogen of the tropics. A transgene was introduced via CRISPR-Cas-assisted homology-directed repair into one of the GSHs in the egg of the parasite. Gene editing efficiencies of 24% and transgene-encoded fluorescence of 75% of gene-edited schistosome eggs were observed. The approach advances functional genomics for schistosomes by providing a tractable path for generating transgenics using homology-directed, repair-catalyzed transgene insertion. We also suggest that this work will serve as a roadmap for the development of similar approaches in helminths more broadly.
鉴定和描述基因组安全港位点（GSH）可以促进转基因的稳定表达，同时最小化对宿主细胞基因组的干扰。我们结合计算基因组注释和染色质结构分析，预测了热带地区主要传染病原体——曼氏血吸虫中的四个 GSH 的位置。通过 CRISPR-Cas 辅助同源定向修复，将转基因导入寄生虫卵中的一个 GSH 中。观察到基因编辑效率为 24%，经基因编辑的血吸虫卵中转基因编码的荧光为 75%。该方法通过同源定向修复催化转基因插入，为曼氏血吸虫提供了一种可行的转基因生成途径，从而推进了血吸虫的功能基因组学研究。我们还认为，这项工作将为更广泛地开发类似的寄生虫方法提供路线图。
