Pulsed SILAC as a Approach for miRNA Targets Identification in Cell Culture

Methods Mol Biol. 2017:1546:149-159. doi: 10.1007/978-1-4939-6730-8_11.

Abstract

Pulsed stable isotope labeling by amino acids in cell culture (pSILAC) comprises a variation of the classical SILAC proteomic methodology that enables the identification of short-term proteomic responses such as those elicited by micro RNAs (miRNAs). Here, we describe a detailed pSILAC protocol for global identification and quantification of protein translation alterations induced by a miRNA using 3T3-L1 pre-adipocytes as a model system.

Keywords: Cell culture; MicroRNAs; Proteomics; Pulsed stable isotope labeling by amino acids; Targets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Cell Culture Techniques
  • Cells, Cultured
  • Gene Expression Regulation*
  • Humans
  • Isotope Labeling
  • MicroRNAs / genetics*
  • Protein Biosynthesis
  • Proteomics / methods*
  • RNA Interference*
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism*
  • Software
  • Workflow

Substances

  • Amino Acids
  • MicroRNAs
  • RNA, Messenger