Knockdown of Human AMPK Using the CRISPR/Cas9 Genome-Editing System

Methods Mol Biol. 2018:1732:171-194. doi: 10.1007/978-1-4939-7598-3_11.

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 α1/α2 catalytic subunits in human cell lines by the CRISPR/Cas9 technology, a recent breakthrough technique for genome editing.

Keywords: AMPK; AMPKα1; AMPKα2; CRISPR/Cas9; PRKAA1; PRKAA2, AML, Caco2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / genetics*
  • CRISPR-Cas Systems / genetics*
  • Gene Editing / methods*
  • Gene Knockdown Techniques / methods*
  • Genetic Vectors / genetics
  • HEK293 Cells
  • Humans
  • Lentivirus / genetics
  • Protein Subunits / genetics
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Signal Transduction / genetics
  • Transfection / methods

Substances

  • Protein Subunits
  • RNA, Guide, CRISPR-Cas Systems
  • PRKAA2 protein, human
  • AMP-Activated Protein Kinases
  • PRKAA1 protein, human