Generation of an shRNAi expression library against the whole human transcripts.
RNA interference is a phenomenon in which expression of an individual gene can be specifically silenced by introducing a double-stranded RNA, homologous to the gene, and is receiving attention as a powerful tool for reverse genetics in the post-genome era. Throughout our current research to generate an siRNA expression library for the whole human genome, we face many technical difficulties. We present here the strategies for overcoming some of the difficulties, including the development of genetically stable and highly active siRNA expression vectors, the selection procedure of the favorable target sites, and the efficient and low cost procedure for constructing an siRNA expression library. Furthermore, we demonstrate that the screening using the constructed siRNA-expression library indeed works, by evaluating siRNA-expression library against apoptosis-related genes.
Miyagishi
Makoto
M
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Hongo, Japan.
Matsumoto
Sahohime
S
Taira
Kazunari
K
D000465:Algorithms; D017209:Apoptosis / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors; D015894:Genome, Human; D006367:HeLa Cells; D006801:Humans; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism
D034741:RNA, Small Interfering