Aptazyme-based riboswitches and logic gates in mammalian cells

Yoko Nomura, Yohei Yokobayashi

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

This chapter describes a screening strategy to engineer synthetic riboswitches that can chemically regulate gene expression in mammalian cells. Riboswitch libraries are constructed by randomizing the key nucleotides that couple the molecular recognition function of an aptamer with the self-cleavage activity of a ribozyme. The allosteric ribozyme (aptazyme) candidates are cloned in the 3′ untranslated region (UTR) of a reporter gene mRNA. The plasmid-encoded riboswitch candidates are transfected into a mammalian cell line to screen for the desired riboswitch function. Furthermore, multiple aptazymes can be cloned into the 3′ UTR of a desired gene to obtain a logic gate response to multiple chemical signals. This screening strategy complements other methods to engineer robust mammalian riboswitches to control gene expression.

Original languageEnglish (US)
Pages (from-to)141-148
Number of pages8
JournalMethods in Molecular Biology
Volume1316
DOIs
StatePublished - May 16 2015
Externally publishedYes

Keywords

  • Aptamer
  • Aptazyme
  • Gene regulation
  • Riboswitch
  • Ribozyme
  • RNA engineering

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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