Engineering protein-responsive mRNA switch in mammalian cells

Methods Mol Biol. 2014;1111:183-96. doi: 10.1007/978-1-62703-755-6_13.

Abstract

Engineering of translation provides an alternative regulatory layer for controlling transgene expression in addition to transcriptional regulation. Synthetic mRNA switches that modulate translation of a target gene of interest in response to an intracellular protein could be a key regulator to construct a genetic circuit. Insertion of a protein binding RNA sequence in the 5' UTR of mRNA would allow for the generation of a protein-responsive RNA switch. Here we describe the design principle of the switch and methods for tuning and analyzing its translational activity in mammalian cells.

MeSH terms

  • 5' Untranslated Regions
  • Animals
  • Apoptosis
  • Base Sequence
  • Cell Culture Techniques / methods
  • Cell Line
  • Flow Cytometry / methods
  • Genes, Reporter
  • Genetic Engineering / methods*
  • Humans
  • Plasmids / genetics
  • Protein Biosynthesis
  • Proteins / genetics
  • Proteins / metabolism*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Riboswitch*
  • Transfection

Substances

  • 5' Untranslated Regions
  • Proteins
  • RNA, Messenger
  • Riboswitch