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J Biomol Tech. 2007 February; 18(1): 86.
PMCID: PMC2291874

SP22 A Lentiviral Vector-Based RNAi Library Screen to Identify Genes that Modulate Cellular Response to Paclitaxel


The delivery of short hairpin RNA (shRNA) through use of lentiviral vectors is proving to be a powerful means to mediate gene-specific RNA interference in mammalian cells. The RNAi Consortium has created a library of 150,000 shRNA clones designed to target 15,000 human and 15,000 mouse genes. Through the utilization of a recombinant lentivirus delivery system, this library is proving to be useful in targeting cell lines that are not amenable to small interfering RNA transfection, and in facilitating screens and experiments that require long-term knockdown that cannot be achieved by synthetic siRNA. After validation of this library and demonstration of its applicability in a broad array of cell types, we conducted a screen using a panel of shRNAs targeting tumor-suppressor genes. We examined the effect of knocking out each of these tumor suppressors in a non-small-cell lung cancer cell line prior to treatment with paclitaxel. This screen was effective in identification of genes involved in increasing sensitization or resistance to this chemotherapeutic. We believe that these results validate the usefulness of shRNA in one screening process designed to identify potential therapeutic targets and biomarkers.

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