Functional genomic analysis of cell division by endoribonuclease-prepared siRNAs.
The emergence of RNA interference (RNAi) technology has revolutionized functional genomic analyses in cell biology, including the study of cell division. In particular, the introduction of short interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) has facilitated loss-of-function studies in mammalian cell lines. We have pioneered the rapid and cost-efficient generation of libraries of endoribonuclease-prepared short interfering RNAs (esiRNAs) for large-scale genetic screens in mammalian tissue culture cells. Our first pilot screen of 5305 genes in human HeLa cells identified novel genes required for cell division, establishing esiRNA as a sophisticated method for loss-of-function screens. Future genome-wide studies with esiRNA, using image-based assays and video microscopy, will help provide novel insights on the molecular mechanisms that govern cell division in mammalian cells.
Kittler
Ralf
R
Max-Planck-Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
Buchholz
Frank
F
D000818:Animals; D002453:Cell Cycle; D002455:Cell Division; D004722:Endoribonucleases / Q000378:metabolism*; D015723:Gene Library; D020868:Gene Silencing; D005821:Genetic Techniques; D016678:Genome; D023281:Genomics / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D018715:Microscopy, Video; D008957:Models, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering
D034741:RNA, Small Interfering; D004722:Endoribonucleases