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A Screen of shRNAs Targeting Tumor Suppressor Genes to Identify Factors Involved in Paclitaxel Sensitivity

> Features of the pLKO.1-Puro vector allow for transient or stable transfection of the shRNA as well as production of lentiviral particles. Unlike adenovirus or murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the specific shRNA construct into differentiated and non-dividing cells, overcoming low transfection and integration difficulties when using these cell lines. pLKO.1 allows for long-term knockdown and phenotypic observation and transduction of difficult or sensitive cell lines.



Figure 2. Screening Tumor Suppressor Knockdown Cells for Paclitaxel Sensitivity.
The MISSION TRC Human Tumor Suppressor Gene Family Set (Cat. No. SH0531) was used to screen for genes that could enhance cell sensitivity to the widely used cancer therapy drug, Paclitaxel. Lentiviral particles were used to transduce cells in 96-well plates and knock down the corresponding tumor suppressor gene expression. Cells were split into two samples. One sample set was treated with Paclitaxel and the second set received medium only. Results were normalized to pLKO.1-puro empty vector control virus (Cat. No. SHC001V). Outliers were observed that either conferred more resistance or sensitivity to Paclitaxel treatment. The red data point represents a positive control for increased sensitivity to Paclitaxel.


Figure 3. Multiple MISSIONTM shRNA Clones Per Target Produce the Same Phenotype.
Multiple clones for four genes conferred increased cell sensitivity to drug treatment. These genes have the potential to be prognostic markers in chemotherapeutic responses.


Similar article originally printed in Genetic Engineering News , Jan. 15,
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