Short-hairpin RNA expressed from polymerase III promoters mediates RNA interference in mosquito cells

D. S. Konet, Joseph S Anderson, J. Piper, R. Akkina, E. Suchman, J. Carlson

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Putative U6snRNA polymerase III (PolIII) promoters were cloned from the Anopheles gambiae and Aedes aegypti genomes. The PolIII promoters were tested for their ability to express short-hairpin RNA (shRNA) targeted to firefly luciferase and to mediate RNA interference (RNAi) knockdown of a co-transfected luciferase reporter gene vector in AG-55 Anopheles gambiae and ATC-10 Aedes aegypti cells. Promoters capable of silencing expression of the co-transfected luciferase plasmid by up to 95% in AG-55 cells and up to 75% in ATC-10 cells were identified. RNase protection experiments allowed detection of the 19 nt luciferase short-interfering RNA (siRNA) in transfected cells. These findings indicate that mosquito U6snRNA gene promoters can be used for production of shRNA to induce the RNAi response in mosquito cells.

Original languageEnglish (US)
Pages (from-to)199-206
Number of pages8
JournalInsect Molecular Biology
Volume16
Issue number2
DOIs
StatePublished - Apr 2007
Externally publishedYes

Keywords

  • Aedes aegypti
  • Anopheles gambiae
  • RNA interference
  • shRNA
  • U6 promoter

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

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