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

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.

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Work Title Gene Editing in the Chagas Disease Vector Rhodnius prolixus by Cas9-Mediated ReMOT Control
Access
Open Access
Creators
  1. Leonardo Lima
  2. Mateus Berni
  3. Jamile Mota
  4. Daniel Bressan
  5. Alison Julio
  6. Robson Cavalcante
  7. Vanessa Macias
  8. Zhiqian Li
  9. Jason L. Rasgon
  10. Ethan Bier
  11. Helena Araujo
Keyword
  1. CRISPR
  2. CRISPR/Cas9 technology
  3. Genetic Engineering, Biotechnology, and Regenerative medicine
  4. Genome editing
License In Copyright (Rights Reserved)
Work Type Article
Publisher
  1. CRISPR Journal
Publication Date April 16, 2024
Publisher Identifier (DOI)
  1. https://doi.org/10.1089/crispr.2023.0076
Deposited August 19, 2024

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Version 1
published

  • Created
  • Added 724609.pdf
  • Added Creator Leonardo Lima
  • Added Creator Mateus Berni
  • Added Creator Jamile Mota
  • Added Creator Daniel Bressan
  • Added Creator Alison Julio
  • Added Creator Robson Cavalcante
  • Added Creator Vanessa Macias
  • Added Creator Zhiqian Li
  • Added Creator Jason L. Rasgon
  • Added Creator Ethan Bier
  • Added Creator Helena Araujo
  • Published
  • Updated
  • Updated Keyword, Publication Date Show Changes
    Keyword
    • CRISPR, CRISPR/Cas9 technology, Genetic Engineering, Biotechnology, and Regenerative medicine, Genome editing
    Publication Date
    • 2024-04-01
    • 2024-04-16