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Short‐hairpin RNA expressed from polymerase III promoters mediates RNA interference in mosquito cells
Author(s) -
Konet D. S.,
Anderson J.,
Piper J.,
Akkina R.,
Suchman E.,
Carlson J.
Publication year - 2007
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.2006.00714.x
Subject(s) - rna interference , small hairpin rna , biology , promoter , microbiology and biotechnology , luciferase , gene knockdown , transfection , rna silencing , small interfering rna , rna polymerase iii , gene silencing , rna , gene , rna polymerase , gene expression , genetics
Putative U6snRNA polymerase III ( Pol III) promoters were cloned from the Anopheles gambiae and Aedes aegypti genomes. The Pol III 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.