Abstract
Exposure to DNA-damaging agents invokes biological responses necessary for damage recovery and cell survival. Despite the presence of intact DNA repair pathways, lack of certain other biological pathways has been shown to sensitize cells to DNA-damaging agents’ exposure. It is likely that following DNA damage a complex interplay between DNA repair pathways and other biological pathways might be required to ensure cell survival. In this chapter, we describe a high-throughput method for the identification of genes essential for cell survival following DNA damage by using a cell-based assay to measure viability in combination with an RNA interference-based genome-wide screening experiment.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mathey-Prevot B, Perrimon N (2006) Drosophila genome-wide RNAi screens: are they delivering the promise? Cold Spring Harb Symp Quant Biol 71:141–148
Ravi D, Wiles AM, Bhavani S, Ruan J, Leder P, Bishop AJ (2009) A network of conserved damage survival pathways revealed by a genomic RNAi screen. PLoS Genet 5:e1000527
Boutros M, Kiger AA, Armknecht S, Kerr K, Hild M, Koch B, Haas SA, Paro R, Perrimon N (2004) Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303:832–835
Ramadan N, Flockhart I, Booker M, Perrimon N, Mathey-Prevot B (2007) Design and implementation of high-throughput RNAi screens in cultured Drosophila cells. Nat Protoc 2:2245–2264
Wiles AM, Ravi D, Bhavani S, Bishop AJ (2008) An analysis of normalization methods for Drosophila RNAi genomic screens and development of a robust validation scheme. J Biomol Screen 13:777–784
Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP (2003) Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res 31:e15
Acknowledgement
This work was supported by NIH grants K22-ES12264 and R15ES019128 as well as a Voelcker Fund Young Investigator Award from the Max and Minnie Tomerlin Voelcker Fund to AJRB. We also thank Aparna Gorthi for her constructive criticism and editing of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Ravi, D., Bishop, A.J.R. (2012). Identification of Genes Required for Damage Survival Using a Cell-Based RNAi Screen Against the Drosophila Genome. In: Bjergbæk, L. (eds) DNA Repair Protocols. Methods in Molecular Biology, vol 920. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-998-3_2
Download citation
DOI: https://doi.org/10.1007/978-1-61779-998-3_2
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-997-6
Online ISBN: 978-1-61779-998-3
eBook Packages: Springer Protocols