@inbook{055f5b7fe8124d30a71ec9dea94eb530,
title = "RNA interference screening methods to identify proliferation determinants and mechanisms of resistance to immune attack",
abstract = "We have used RNA interference (RNAi) screening technology to reveal unknown components of biological signaling pathways including survival mechanisms of estrogen-independent breast cancer cell growth and cancer cell resistance to immune attack. In this chapter, a detailed protocol describing the use of RNAi screening to identify factors important for the proliferation of estrogen-independent MCF7 breast cancer cells will be described. Resistance to therapies that target the estrogen pathway remains a challenge in the treatment of estrogen receptor-positive breast cancer. To address this challenge, small interfering-RNA (siRNA)-based libraries targeting an estrogen receptor (ER)- and aromatase-centered network, including 631 genes relevant to estrogen signaling, was designed and constructed for RNAi screening. This protocol will include the following parts: (1) selection of RNAi transfection reagent for specific cells; (2) optimization of RNAi screening conditions using Z′-factor; (3) procedure of ER-network gene siRNA library screening using automated machines under optimized experimental conditions; and (4) method of analysis for RNAi screening data to identify specific determinants important for cell proliferation. 46 genes were found to be selectively required for the survival of estrogen-independent MCF7-derived cells.",
keywords = "Breast cancer, CyBio automatic dispenser, Estrogen receptor (ER), Immunotherapy, Multidrop Combi-nL reagent dispenser, RNA interference (RNAi), Screening, WellMate microplate dispenser, Z′-factor, Humans, RNA, Small Interfering/genetics, Gene Expression Regulation, Neoplastic, MCF-7 Cells, RNA Interference, Cell Line, Tumor, Cell Proliferation/genetics, Female, Breast Neoplasms/genetics, Receptors, Estrogen/genetics",
author = "Zhang, \{Yong Wei\} and Nasto, \{Rochelle E.\} and Jablonski, \{Sandra A.\} and Serebriiskii, \{Ilya G.\} and Rishi Surana and Joseph Murray and Michael Johnson and Riggins, \{Rebecca B.\} and Robert Clarke and Golemis, \{Erica A.\} and Weiner, \{Louis M.\}",
note = "Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
doi = "10.1016/bs.mie.2019.06.002",
language = "English",
isbn = "9780128206676",
volume = "636",
series = "Methods in enzymology",
publisher = "Academic Press Inc.",
pages = "299--322",
editor = "Lorenzo Galluzzi and Nils-Petter Rudqvist",
booktitle = "Tumor Immunology and Immunotherapy - Integrated Methods Part B",
address = "United States",
}