Program in Cell and Developmental Biology, Institute for Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, Rio de Janeiro, Brazil.
Department of Biological Sciences, University of North Texas, Denton, Texas, USA.
CRISPR J. 2024 Apr;7(2):88-99. doi: 10.1089/crispr.2023.0076. Epub 2024 Apr 1.
is currently the model vector of choice for studying Chagas disease transmission, a debilitating disease caused by parasites. However, transgenesis and gene editing protocols to advance the field are still lacking. Here, we tested protocols for the maternal delivery of CRISPR-Cas9 (clustered regularly spaced palindromic repeats/Cas-9 associated) elements to developing oocytes and strategies for the identification of insertions and deletions (indels) in target loci of resulting gene-edited generation zero (G0) nymphs. We demonstrate successful gene editing of the eye color markers and , and the cuticle color marker with highest effectiveness obtained using Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) with the ovary-targeting BtKV ligand. These results provide proof of concepts for generating somatic mutations in and potentially for generating germ line-edited lines in triatomines, laying the foundation for gene editing protocols that could lead to the development of novel control strategies for vectors of Chagas disease.
目前，它是研究恰加斯病传播的首选模型载体，恰加斯病是由寄生虫引起的一种使人虚弱的疾病。然而，推进这一领域的转基因和基因编辑技术仍很缺乏。在这里，我们测试了将 CRISPR-Cas9（成簇规律间隔短回文重复序列/ Cas9 相关）元件通过母体传递到发育中的卵母细胞的方案，以及鉴定目标基因编辑零代（G0）若虫靶基因座中插入和缺失（indels）的策略。我们成功地编辑了眼睛颜色标记 和 ，以及表皮颜色标记 ，使用靶向卵巢的 BtKV 配体的受体介导卵巢转导（ReMOT Control），获得了最高的编辑效率。这些结果为在 和 中产生体突变提供了概念验证，也为在三锥虫中产生生殖系编辑系奠定了基础，为基因编辑技术奠定了基础，可能会开发出针对恰加斯病传播媒介的新型控制策略。
