Genome editing efficiency of four Drosophila suzukii endogenous U6 promoters

Insect Mol Biol. 2021 Aug;30(4):420-426. doi: 10.1111/imb.12707. Epub 2021 May 24.

Abstract

The invasive spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) has caused serious economic losses to the fruit industry. The conventional control methods have many limitations and genetic engineering technologies such as CRISPR/Cas9-mediated gene drive are promising approaches. In the CRISPR/Cas9 system, the transcriptional regulatory elements play an important role in the activities of gRNA. Thus, in order to improve the genome editing efficiency of the CRISPR/Cas9 system in D. suzukii, we cloned and tested four endogenous U6 promoters to drive mutagenesis of the white gene. Our results showed that all the four promoters could be used with variable efficiency. The promoter DsU6-3 had the highest genome editing efficiency among the four DsU6 promoters. Compared with the DsU6-3 promoter, the DmU6:3 promoter showed lower efficiency to drive mutagenesis in D. suzukii. These findings expand the range of promoters available to express gRNAs in D. suzukii, facilitating the basic and applied research on this important pest.

Keywords: CRISPR/Cas9; Drosophila suzukii; U6 promoter; genome editing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Drosophila / genetics*
  • Drosophila Proteins / genetics
  • Gene Editing / methods*
  • Genetic Engineering
  • Insect Control
  • Mutagenesis
  • Promoter Regions, Genetic / genetics*

Substances

  • Drosophila Proteins