Summary
In mammalian cells, apoptotic and anti-apoptotic pathways may be investigated using a variety of biochemical, molecular, and genetic approaches. Retrovirus mediated genetic screens have proven a powerful tool in mapping out the network of players in a number of signaling pathways. We have developed the ERM (for enhanced retroviral mutagen) mutagenesis approach to identify novel players in the growth factor dependent survival pathways. ERM has been shown to be efficient and amenable to genome wide genetic screens in mammalian cells without the need of cDNA library construction. The advantages of the ERM method include regulatable expression, flexible design, and efficiency.
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
Liu, D., Yang, X., and Songyang, Z. (2000). Identification of CISK, a new member of the SGK kinase family that promotes IL-3-dependent survival. Curr Biol 10, 1233–6.
Ding, Z., Liang, J., Lu, Y., Yu, Q., Songyang, Z., Lin, S. Y., and Mills, G. B. (2006). A retrovirus-based protein complementation assay screen reveals functional AKT1-binding partners. Proc Natl Acad Sci USA 103, 15014–9.
Liu, D., Yang, X., Yang, D., and Songyang, Z. (2000). Genetic screens in mammalian cells by enhanced retroviral mutagens. Oncogene 19, 5964–72.
Pear, W. S., Nolan, G. P., Scott, M. L., and Baltimore, D. (1993). Production of high-titer helper-free retroviruses by transient transfection. Proc Natl Acad Sci USA 90, 8392–96.
Morgenstern, J. P., and Land, H. (1990). Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res 18, 3587–96.
Gossen, M., Bonin, A. L., and Bujard, H. (1993). Control of gene activity in higher eukaryotic cells by prokaryotic regulatory elements. Trends Biochem Sci 18, 471–5.
Paulus, W., Baur, I., Boyce, F. M., Breakefield, X. O., and Reeves, S. A. (1996). Self-contained, tetracycline-regulated retroviral vector system for gene delivery to mammalian cells. J Virol 70, 62–7.
Brent, M. R. (2005). Genome annotation past, present, and future: how to define an ORF at each locus. Genome Res 15, 1777–86.
Suzuki, T., Shen, H., Akagi, K., Morse, H. C., Malley, J. D., Naiman, D. Q., Jenkins, N. A., and Copeland, N. G. (2002). New genes involved in cancer identified by retroviral tagging. Nat Genet 32, 166–74.
Acknowledgments
This work is supported by the Welch Foundation (to Z.S.) and NIH grants CA84208 and GM69572. D.L. is supported in part by the American Heart Association. Z.S. is a Leukemia and Lymphoma Society Scholar.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Liu, D., Songyang, Z. (2009). Genetic Mapping of Anti-Apoptosis Pathways in Myeloid Progenitor Cells. In: Erhardt, P., Toth, A. (eds) Apoptosis. Methods in Molecular Biology, vol 559. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-017-5_20
Download citation
DOI: https://doi.org/10.1007/978-1-60327-017-5_20
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-016-8
Online ISBN: 978-1-60327-017-5
eBook Packages: Springer Protocols