title: Short-hairpin RNA expressed from polymerase III promoters mediates RNA interference in mosquito cells.
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.
authors.affiliation: Department of Microbiology Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA.
authors.forename: D S
authors.initials: DS
authors.lastname: Konet
authors.forename: J
authors.initials: J
authors.lastname: Anderson
authors.forename: J
authors.initials: J
authors.lastname: Piper
authors.forename: R
authors.initials: R
authors.lastname: Akkina
authors.forename: E
authors.initials: E
authors.lastname: Suchman
authors.forename: J
authors.initials: J
authors.lastname: Carlson
mesh_terms: D000330:Aedes / Q000235:genetics*; D000818:Animals; D000852:Anopheles / Q000235:genetics*; D001483:Base Sequence; D049409:Luciferases, Firefly / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012320:RNA Polymerase III / Q000235:genetics; D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear; C405486:U6 small nuclear RNA; D049409:Luciferases, Firefly; D012320:RNA Polymerase III