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

Bartlomiej Bartkowiak, Christopher Yan, Arno L Greenleaf

Biochimica et biophysica acta 2015 Sep

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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. Copyright © 2015 Elsevier B.V. All rights reserved.

Citation

Bartlomiej Bartkowiak, Christopher Yan, Arno L Greenleaf. Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas. Biochimica et biophysica acta. 2015 Sep;1849(9):1179-87