Sirna Expression Plasmid Library Construction,

Abstract: RNA interference(RNAi) was a post transcriptional silencing of homologous process in which double-stranded RNA (dsRNA) induces specific transcripts. Systematic silencing of genes on a genome-wide scale functional gene screening using small interference(siRNA) library, which was targeting many genes, provided a powerful research tool in functional genomics. Owing to the expensive cost of obtaining the siRNA by way of chemosynthesis, it was very significant to construct a siRNA library which could be expanded. Aiming at constructing a siRNA library for the specific human genes had been known, the library was constructed by adopting a newly designed siRNA expression cassette through pBUH. It was shown in this study respecting to the reporting genes of GFP and RED, the siRNA of EGFP could be effectively expressed, that resulted in the explicit depression of expression level for EGFP. Furthermore, 300 siRNA have been successfully cloned. Finally, the preparative experiment for the functional screening of cell apoptosis relating to genes using this library was pre-fulfilled. The siRNA library constructed would be an important and available resource for the research of functional genomics. Through the procedure research for constructing the library and the research result respecting to the functional gene screening of cell apoptosis induced by staurosporine (simplified as STS), the siRNA library could be further expanded based on the work above mentioned, which founded the basis of the wide-scale screening.