Skip to main content

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

Publication ,  Journal Article
Bartkowiak, B; Yan, C; Greenleaf, AL
Published in: Biochim Biophys Acta
September 2015

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.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

September 2015

Volume

1849

Issue

9

Start / End Page

1179 / 1187

Location

Netherlands

Related Subject Headings

  • Sequence Homology, Nucleic Acid
  • Molecular Sequence Data
  • Humans
  • Hela Cells
  • HeLa Cells
  • Gene Knockdown Techniques
  • DNA
  • Cyclin-Dependent Kinases
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Base Sequence
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Bartkowiak, B., Yan, C., & Greenleaf, A. L. (2015). Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas. Biochim Biophys Acta, 1849(9), 1179–1187. https://doi.org/10.1016/j.bbagrm.2015.07.010
Bartkowiak, Bartlomiej, Christopher Yan, and Arno L. Greenleaf. “Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas.Biochim Biophys Acta 1849, no. 9 (September 2015): 1179–87. https://doi.org/10.1016/j.bbagrm.2015.07.010.
Bartkowiak B, Yan C, Greenleaf AL. Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas. Biochim Biophys Acta. 2015 Sep;1849(9):1179–87.
Bartkowiak, Bartlomiej, et al. “Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas.Biochim Biophys Acta, vol. 1849, no. 9, Sept. 2015, pp. 1179–87. Pubmed, doi:10.1016/j.bbagrm.2015.07.010.
Bartkowiak B, Yan C, Greenleaf AL. Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas. Biochim Biophys Acta. 2015 Sep;1849(9):1179–1187.

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

September 2015

Volume

1849

Issue

9

Start / End Page

1179 / 1187

Location

Netherlands

Related Subject Headings

  • Sequence Homology, Nucleic Acid
  • Molecular Sequence Data
  • Humans
  • Hela Cells
  • HeLa Cells
  • Gene Knockdown Techniques
  • DNA
  • Cyclin-Dependent Kinases
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Base Sequence