Prime Editing for Precise Genome Engineering in Drosophila

Methods Mol Biol. 2022:2540:113-134. doi: 10.1007/978-1-0716-2541-5_5.

Abstract

Editing the Drosophila genome is incredibly useful for gene functional analysis. However, compared to gene knockouts, precise gene editing is difficult to achieve. Prime editing, a recently described CRISPR/Cas9-based technique, has the potential to make precise editing simpler and faster, and produce less errors than traditional methods. Initially described in mammalian cells, prime editing is functional in Drosophila somatic and germ cells. Here, we outline steps to design, generate, and express prime editing components in transgenic flies. Furthermore, we highlight a crossing scheme to produce edited fly stocks in less than 3 months.

Keywords: CRISPR; Drosophila; Genome engineering; Precise editing; Prime editing; pegRNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • CRISPR-Cas Systems* / genetics
  • Drosophila* / genetics
  • Gene Editing / methods
  • Genome, Insect
  • Mammals / genetics