Send to

Choose Destination
Methods Mol Biol. 2009;545:1-19. doi: 10.1007/978-1-60327-993-2_1.

Knock-in and knock-out: the use of reverse genetics in somatic cells to dissect mitotic pathways.

Author information

Department of Oncology, Cancer Research UK Cambridge Research Institute, University of Cambridge, Cambridge, UK.


Reverse genetic methods, such as homologous gene targeting, have greatly contributed to our understanding of molecular pathways in mitosis, especially in yeast. The chicken B-lymphocyte line, DT40, represents a unique example among vertebrate somatic cells where homologous gene targeting occurs at very high frequency. DT40 cells therefore provide a useful and accessible somatic genetic system for wide-ranging biochemical and cell biological assays. In this chapter, we describe the main principles of homologous gene targeting, the concept of targeting construct design and the detailed experimental protocol of how to achieve successful knockouts. We also mention methods for conditional disruption of essential genes and conclude with specific procedures for the study of mitosis in DT40 cells.

[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center