2008
DOI: 10.1126/science.1149200
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Cancer Proliferation Gene Discovery Through Functional Genomics

Abstract: Retroviral short hairpin RNA (shRNA)-mediated genetic screens in mammalian cells are powerful tools for discovering loss-of-function phenotypes. We describe a highly parallel multiplex methodology for screening large pools of shRNAs using half-hairpin barcodes for microarray deconvolution. We carried out dropout screens for shRNAs that affect cell proliferation and viability in cancer cells and normal cells. We identified many shRNAs to be antiproliferative that target core cellular processes, such as the cell… Show more

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Cited by 352 publications
(351 citation statements)
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References 19 publications
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“…We performed a genome-wide RNAi screen by using a pooled lentiviral shRNA library (15,16) to identify regulators of apoptosis. To help quickly remove off-target effects, we validated the ability to repeat the FASL sensitization phenotype of the top depleted hits by using siRNAs with unique target sequences.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…We performed a genome-wide RNAi screen by using a pooled lentiviral shRNA library (15,16) to identify regulators of apoptosis. To help quickly remove off-target effects, we validated the ability to repeat the FASL sensitization phenotype of the top depleted hits by using siRNAs with unique target sequences.…”
Section: Discussionmentioning
confidence: 99%
“…To identify regulators of death receptor-induced apoptosis, we performed a genome-wide RNAi screen by using the pooledshRNA approach described previously by others (15)(16)(17). This approach led to the identification of 16 hits whose knockdown increases the sensitivity of cells to FASL.…”
mentioning
confidence: 99%
“…Although the identification of synthetic lethal lead compounds through large-scale chemical screening may be hard to come by because of chemical scarcity and large diversity, genetic synthetic lethality screening for gene target identification through siRNAs (Iorns et al, 2007) or constitutive/inducible shRNA libraries (Ngo et al, 2006;Schlabach et al, 2008, respectively) seems more promising. This is due, in part, to the limited number of human genes, and the increased efficiencies of the RNAi tools.…”
Section: Resultsmentioning
confidence: 99%
“…NIH3T3 fibroblast cells were infected with the 63,996 pooled lentiviral mouse shRNA library at a multiplicity of infection (m.o.i.) of one 14,15 . Two days after infection, shRNA-infected cells were selected with puromycin, placed into the upper compartment of a transwell unit and allowed to migrate through the perforated membrane to the lower compartment.…”
Section: Genome-wide Functional Selection Of Cell Migration Regulatorsmentioning
confidence: 99%