Gene Editing in the Chagas Disease Vector Rhodnius prolixus by Cas9-Mediated ReMOT Control

Leonardo Lima, Mateus Berni, Jamile Mota, Daniel Bressan, Alison Julio, Robson Cavalcante, Vanessa Macias, Zhiqian Li, Jason L. Rasgon, Ethan Bier, Helena Araujo

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Rhodnius prolixus is currently the model vector of choice for studying Chagas disease transmission, a debilitating disease caused by Trypanosoma cruzi 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 R. prolixus 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 Rp-scarlet and Rp-white, and the cuticle color marker Rp-yellow, 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 R. prolixus 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.

Original languageEnglish (US)
Pages (from-to)88-99
Number of pages12
JournalCRISPR Journal
Issue number2
StatePublished - Apr 1 2024

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Genetics

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