RNA Interference Screening to Identify Proliferation Determinants in Breast Cancer Cells
Yong-Wei Zhang,
Rochelle Nasto,
Sandra Jablonski,
Ilya Serebriiskii,
Rishi Surana,
Joseph Murray,
Michael Johnson,
Rebecca Riggins,
Robert Clarke,
Erica Golemis,
Louis Weiner
Affiliations
Yong-Wei Zhang
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Rochelle Nasto
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USAProgram in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, USA, School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, PA, USA
Sandra Jablonski
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Ilya Serebriiskii
Program in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, USAKazan Federal University, Kazan, Russia
Rishi Surana
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Joseph Murray
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Michael Johnson
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Rebecca Riggins
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Robert Clarke
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
Erica Golemis
Program in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, USA
Louis Weiner
Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, USA
RNAi screening technology has revealed unknown determinants of various biological signaling pathways in biomedical studies. This protocol provided detailed information about how to use RNAi screening to identify proliferation determinants in breast tumor cells. siRNA-based libraries targeting against Estrogen receptor (ER)-network, including 631 genes relevant to estrogen signaling, was constructed for screening in breast cancer cells. Briefly, reverse transfection of siRNA induced transient gene knockdown in MCF7 cells. First, the transfection reagent for MCF7 cells was selected. Next, the Z’-score assay was used to monitor if screening conditions yielded efficiently. Then, the ER-network siRNA library screening was preceded by automatic machines under optimized experimental conditions.
