Grenier Adrien, Sujobert Pierre, Olivier Séverine, Guermouche Hélène, Mondésir Johanna, Kosmider Olivier, Viollet Benoit, Tamburini Jérôme
U1016, Institut Cochin, Inserm, Paris, France.
UMR8104, CNRS, Paris, France.
Methods Mol Biol. 2018;1732:171-194. doi: 10.1007/978-1-4939-7598-3_11.
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.
AMP 激活的蛋白激酶（AMPK）是一种关键的能量传感器，可调节涉及包括代谢疾病和癌症在内的病理学的信号网络。AMPK 这一日益被认可的作用促使人们付出巨大的研究努力来开发新的药理学 AMPK 激活剂。为了在细胞模型中精确研究 AMPK 的作用以及 AMPK 激活剂的特异性和活性，需要基因 AMPK 失活工具。我们在此报告通过 CRISPR/Cas9 技术（一种用于基因组编辑的最新突破性技术）在人细胞系中对 AMPK α1/α2 催化亚基进行基因失活的方法。
