Short-hairpin RNA expressed from polymerase III promoters mediates RNA interference in mosquito cells
Autor(es): Konet D S,Anderson J,Piper J,Akkina R,Suchman E,Carlson J
Resumo: 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.
Palavras-Chave: RNA interference; shRNA; U6 promoter; Aedes aegypti; Anopheles gambiae
Imprenta: Insect Molecular Biology, v. 16, n. 2, p. 199-206, 2007
Identificador do objeto digital: 10.1111/j.1365-2583.2006.00714.x
Descritores: Aedes aegypti - Cell ; Aedes aegypti - Cytopathology ; Aedes aegypti - Genome ; Aedes aegypti - Molecular Structure ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Proteins ; Aedes aegypti - RNA ; Aedes aegypti - Immunology
Data de publicação: 2007
