CRISPR-Mediated Genome Engineering in Aedes aegypti

Ruichen Sun, Ming Li, Conor J. McMeniman, Omar S. Akbari

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

CRISPR-mediated genome engineering technologies have been adapted to a wide variety of organisms with high efficiency and specificity. The yellow fever mosquito, Aedes aegypti, is one such organism. It is also responsible for transmitting a wide variety of deadly viruses including Dengue, Zika, Yellow fever, and Chikungunya. The key to successful CRISPR-mediated gene editing applications is the delivery of both Cas9 ribonuclease and single-guide RNA (sgRNA) to the nucleus of desired cells. Various methods have been developed for supplying the Cas9 endonuclease, sgRNA, and donor DNA to Ae. aegypti. In this chapter, we focus on methods of direct embryo delivery of editing components, presenting detailed step-by-step CRISPR/Cas9-based genome-editing protocols for inducing desired heritable edits in mosquitoes as well as insights into successful application of these protocols. We also highlight potential opportunities for customizing these protocols to manipulate the mosquito genome for innovative in vivo gene function studies.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages23-51
Number of pages29
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2509
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Aedes aegypti
  • CRISPR/Cas9
  • Gene editing
  • HDR
  • sgRNA

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

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