shRNA-mediated RNA interference as a tool for genetic synthetic lethality screening in mouse embryo fibroblasts.
Previously, we demonstrated the establishment of synthetic lethality screening in cultured somatic human cells, or mouse embryo fibroblasts (MEFs), for chemicals or mutant genes synergistically lethal with a mutated gene of interest. Here, we show in MEFs that the usage of RNA interference-based genetic suppressor elements encoding short hairpin RNAs (shRNAs) enables for genetic synthetic lethality screening at a frequency much higher than that achieved before with short truncated sense and antisense RNAs. These findings open up the possibility of using in mammalian cells genome-wide shRNA libraries for genetic synthetic lethality screening at the multi-gene level.
Einav
Yulia
Y
Department of Biochemistry, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv 69978, Israel.
Agami
Reuven
R
Canaani
Dan
D
D000818:Animals; D019731:Carbon-Nitrogen Ligases / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D005804:Genes, Lethal / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D023281:Genomics / Q000379:methods*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011687:Purines / Q000096:biosynthesis; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*
D011687:Purines; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019731:Carbon-Nitrogen Ligases; C020828:GMP synthase (glutamine-hydrolyzing); C030985:purine