RNA interference (RNAi) mediated by short hairpin-RNA (shRNA) expressing plasmids can induce specific and long-term knockdown of specific mRNAs in eukaryotic cells. To develop a vector-based RNAi model for @entity380 , the schistosome U6 gene promoter was employed to drive expression of shRNA targeting reporter firefly luciferase. An upstream region of a U6 gene predicted to contain the promoter was amplified from genomic DNA of @entity383 . A shRNA construct driven by the predicted U6 promoter targeting luciferase was assembled and cloned into plasmid pXL-Bac II, the construct termed pXL-BacII_SmU6-shLuc. Luciferase expression in transgenic @entity1492 HT-1080 cells was significantly reduced 96 h following transduction with plasmid pXL-BacII_SmU6-shLuc, which encodes luciferase mRNA-specific shRNA. In a similar fashion, schistosomules of @entity383 were transformed with the SmU6-shLuc or control constructs. Firefly luciferase mRNA was introduced into transformed schistosomules after which luciferase activity was analyzed. Significantly less activity was present in schistosomules transfected with pXL-BacII_SmU6-shLuc compared with controls. The findings revealed that the putative @entity383 U6 gene promoter of 270 bp in length was active in @entity1 cells and schistosomes. Given that the U6 gene promoter drove expression of shRNA from an episome, the findings also indicate the potential of this putative RNA polymerase III dependent promoter as a component regulatory element in vector-based RNAi for functional genomics of schistosomes.

@entity380 U6 gene promoter-driven short hairpin RNA induces RNA interference in XXXX @entity1492 cells and schistosomules.

@entity383 :: ('99802', 'Species') :: ['S. mansoni'] @entity1 :: ('9606', 'Species') :: ['human'] @entity380 :: ('6183', 'Species') :: ['Schistosoma mansoni'] @entity1492 :: ('MESH:D005354', 'Disease') :: ['fibrosarcoma']
@entity1 :: (9606,Species) :: ['human']
