Knockdown of human AMPK using the CRISPR-Cas9 genome-editing system

TL;DR

This paper presents a method using CRISPR-Cas9 to genetically inactivate AMPK in human cell lines, enabling precise study of its role in metabolism and disease.

Contribution

It introduces a novel CRISPR-Cas9 based approach for specific knockout of AMPK subunits in human cells, facilitating functional studies.

Findings

Successful knockout of AMPK $\\alpha1/ \\alpha2$ in human cell lines.

Enables detailed investigation of AMPK's role in cellular metabolism.

Provides a tool for testing AMPK-targeting drugs.

Abstract

AMP activated protein kinase (AMPK) is a critical energy sensor, regulating signaling networks involved in pathology including metabolic diseases and cancer. This increasingly recognized role of AMPK has prompted tremendous research efforts to develop new pharmacological AMPK activators. To precisely study the role of AMPK, and the specificity and activity of AMPK activators in cellular models, genetic AMPK inactivating tools are required. We report here methods for genetic inactivation of AMPK $\alpha1/ \alpha2$ catalytic subunits in human cell lines by the CRISPR/Cas9 technology, a recent breakthrough technique for genome editing.