Related Experiment Video
Updated: Apr 6, 2026
Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
Published on: January 3, 2015
Engineering an analog-sensitive CDK12 cell line using CRISPR/Cas
Bartlomiej Bartkowiak1, Christopher Yan1, Arno L Greenleaf1
1Department of Biochemistry, Duke University Medical Center, United States.
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.
Insights
Researchers engineered an analog-sensitive CDK12 cell line to study its function. This tool helps understand CDK12
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- CDK12, a kinase regulating RNA Polymerase II, is linked to tumor suppression and DNA damage response.
- Understanding CDK12 function is hindered by the lack of specific inhibitors and challenges with RNAi.
- New tools are needed to investigate CDK12's precise molecular roles.
Purpose of the Study:
- To develop a novel tool for studying CDK12 function.
- To enable selective and reversible inhibition of CDK12 activity.
Main Methods:
- Utilized CRISPR/Cas gene editing to create an analog-sensitive CDK12 (asCDK12) mammalian cell line.
- Developed a system for inducible inhibition of CDK12 using a cell-permeable adenine analog.
Main Results:
- Inhibition of asCDK12 led to altered CTD phosphorylation patterns.
- CDK12 inhibition resulted in a significant arrest of cellular proliferation.
Conclusions:
- The developed asCDK12 cell line is a powerful new tool for dissecting CDK12 function.
- This system facilitates future research into CDK12's roles in cellular processes and disease.
Related Concept Videos
01:59CRISPR
01:28CRISPR/Cas9 Genome Editing