The Verification Of Alternative Splicing And Preliminary Study Of Mouse Gene Sidt2

RNA interference(RNAi)has been shown to spread from cell to cell and from parent to progeny.This phenomenon was named systemic RNAi.The gene sid-1 is responsible for the spreading of RNAi between cells.It appears to be a mediator of dsRNA uptake and is necessary for systemic RNAi in C.elegans.The homology family of sid1 in human,mouse and rat includes two members-sidt1 and sidt2.We carried out a preliminary study on mouse sidt2.The mouse gene sidt2 includes two sequences,BC051101 and BC023957, with a 63bp difference in length within their coding region.So we named them sidt2(1)and sidt2(s).We analysed the expression profile of sidt2(1/s)in mouse by RT-PCR.And we blasted the mRNA sequences in gene bank with mouse genome sequence and distinguished the genome location of sidt2 with PCR.Then we constructed an alternative splicing reporter vector and verified the alternative splicing of mouse sidt2.Moreover,we silenced the splicing factors,CLK2, SNRPB and hnRNP A1,and detected the splicing of sidt2 by RT-PCR.It appears that these factors can't change the splicing of sidt2.But hnRNP A1 seems to play a role in regulating the expression of some genes.Both structure prediction and subcellular location of mouse SIDT2(L/S) show that they are multispan transmembrane proteins and located at the membrane system of cells.Because of the high homology of sidt2(1)gene between mouse and rat,we isolated primary liver cells of rat and incubated them with FAM-siRNA. We found that the rat liver cells could uptake the siRNA without the presence of lipofectamine,which indicated that sidt2(1)may help the siRNA to enter the liver cells.