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

ABSTRACT: 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.

SUBMITTER: Bartkowiak B

PROVIDER: S-EPMC4556607 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature

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

Bartkowiak Bartlomiej B Yan Christopher C Greenleaf Arno L AL

Biochimica et biophysica acta 20150717 9

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 additi ...[more]

PMID: 26189575

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