RNA Scaffolds pp 141-148 | Cite as

Aptazyme-Based Riboswitches and Logic Gates in Mammalian Cells

Part of the Methods in Molecular Biology book series (MIMB, volume 1316)

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.

Key words

Riboswitch Aptamer Ribozyme Aptazyme Gene regulation RNA engineering 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Biomedical EngineeringUniversity of California, DavisDavisUSA

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