McCurrach, M. E., Lowe, S. W. (2001) Methods for studying pro- and antiapoptotic genes in nonimmortal cells. In: Methods in Cell Biology. Methods in Cell Biology, 66 . Academic Press Inc, San Diego, pp. 197-227. ISBN 0091-679X
Abstract
This chapter describes procedures used in studying gene function in early passage mouse embryo fibroblasts or other nonimmortalized cell types. The approach discussed in the chapter combines high-titer retroviral vectors with various null mutant cells to assess combinations of genes in a defined genetic background. A typical experiment involves four stages: (1) isolation of fibroblasts from normal and/or knockout mice; (2) generation of a retroviral vector; (3) infection of cells and selection for infected populations; and (4) analysis of cell death, transformation. This approach helps mix and match dominant and recessive activities to build model “tumor” cells from normal counterparts without clonal expansion in culture. As a result, one can have strict control over genetic background, thereby avoiding complications arising from unknown mutations that accumulate in immortal or tumor-derived lines. Although the power of this approach is in its well-defined genetic nature, it is important to recognize that these systems are necessarily artificial. Therefore, it is recommended that, having understood pro- or antiapoptotic gene function in normal cells, one tests prediction of these systems in animal models or by using patient material.
| Item Type: | Book Section |
|---|---|
| Uncontrolled Keywords: | TITER RETROVIRAL VECTORS P53-DEPENDENT APOPTOSIS BONE-MARROW STEM-CELLS THERAPY P53 TRANSFORMATION TRANSDUCTION FIBROBLASTS EFFICIENT |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > apoptosis bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function |
| CSHL Authors: | |
| Communities: | CSHL labs > Lowe lab |
| Depositing User: | Matt Covey |
| Date: | 2001 |
| Date Deposited: | 22 Jan 2014 20:23 |
| Last Modified: | 22 Jan 2014 20:23 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29212 |
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