Use of DT40 conditional-knockout cell lines to study chromosomal passenger protein function

Abstract

The CPC [chromosomal passenger complex; INCENP (inner centromere protein), Aurora B kinase, survivin and borealin] is implicated in many mitotic processes. In the present paper we describe how we generated DT40 conditional-knockout cell lines for incenp1 and survivin1 to better understand the role of these CPC subunits in the control of Aurora B kinase activity. These lines enabled us to reassess current knowledge of survivin function and to show that INCENP acts as a rheostat for Aurora B activity.

Aurora B Overexpression Causes Aneuploidy and p21Cip1 Repression during Tumor Development

“…Here, we have taken advantage of a robust system to modulate the expression of endogenous genes in mammals by using a pro- moter-hijack strategy previously used to modulate gene expression in chicken DT40 cells (27,41). Replacement of endogenous promoter regions by tetracycline-responsive elements results in robust control of endogenous gene expression both in vitro and in vivo, eliminating the need to express exogenous genes ( Fig.…”