Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas.

Published

Journal Article

The RNA Polymerase II C-terminal domain (CTD) kinase CDK12 has been implicated as a tumor suppressor and regulator of DNA damage response genes. Although much has been learned about CDK12 and its activity, due to the lack of a specific inhibitor and the complications posed by long term RNAi depletion, much is still unknown about the particulars of CDK12 function. Therefore gaining a better understanding of CDK12's roles at the molecular level will be challenging without the development of additional tools. In order to address these issues we have used the CRISPR/Cas gene engineering system to create a mammalian cell line in which the only functional copy of CDK12 is selectively inhibitable by a cell-permeable adenine analog (analog-sensitive CDK12). Inhibition of CDK12 results in a perturbation of the phosphorylation patterns on the CTD and an arrest in cellular proliferation. This cell line should serve as a powerful tool for future studies.

Full Text

Duke Authors

Cited Authors

  • Bartkowiak, B; Yan, C; Greenleaf, AL

Published Date

  • September 2015

Published In

Volume / Issue

  • 1849 / 9

Start / End Page

  • 1179 - 1187

PubMed ID

  • 26189575

Pubmed Central ID

  • 26189575

International Standard Serial Number (ISSN)

  • 0006-3002

Digital Object Identifier (DOI)

  • 10.1016/j.bbagrm.2015.07.010

Language

  • eng

Conference Location

  • Netherlands