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Updated: Jun 2, 2026
08:35
Inducible and Reversible Dominant-negative (DN) Protein Inhibition
Published on: January 7, 2019
Cell-type specific expression of a dominant negative PKA mutation in mice
Brandon S Willis1, Colleen M Niswender, Thomas Su
1Department of Pharmacology, University of Washington, Seattle, Washington, United States of America.
Plos One
|May 3, 2011
Summary
Researchers developed a novel mouse model to control protein kinase A (PKA) activity using the Cre-loxP system. This tool enables targeted inhibition of PKA, aiding in the study of its physiological roles.
Area of Science:
- Molecular Biology
- Genetics
- Physiology
Background:
- Protein kinase A (PKA) is a crucial enzyme regulating numerous cellular processes.
- Understanding PKA's specific roles in different cell types is challenging due to its widespread activity.
Purpose of the Study:
- To develop a conditional mouse model for inducible PKA inhibition in specific cell types.
- To investigate the physiological consequences of PKA inhibition in vivo.
Main Methods:
- Utilized the Cre recombinase/loxP system to generate a mouse line with a dominant-negative RIαB allele.
- The mutant allele, encoding a substituted RIα regulatory subunit, is expressed upon Cre-mediated excision of a floxed neomycin cassette.
- Assessed PKA activity and gene expression in embryonic stem cells and hepatocytes.
Main Results:
- Cre-mediated activation of the RIαB allele successfully reduced PKA activity in embryonic stem cells and hepatocytes.
- Inhibition of PKA in hepatocytes did not disrupt fasting-induced gene expression but enhanced glucose disposal.
- Demonstrated the feasibility of targeted PKA inhibition for physiological studies.
Conclusions:
- The developed mouse line provides a powerful, cell-type-specific tool for dissecting PKA function.
- Conditional PKA inhibition can modulate metabolic processes like glucose disposal without broadly affecting gene expression.
- This system offers new avenues for studying PKA's role in various physiological contexts.
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