Rational design of aptazyme riboswitches for efficient control of gene expression in mammalian cells
- The Scripps Research Institute, United States
- Johns Hopkins School of Medicine, United States
- University of Massachusetts Medical School, United States
Abstract
Efforts to control mammalian gene expression with ligand-responsive riboswitches have been hindered by lack of a general method for generating efficient switches in mammalian systems. Here we describe a rational-design approach that enables rapid development of efficient cis-acting aptazyme riboswitches. We identified communication-module characteristics associated with aptazyme functionality through analysis of a 32-aptazyme test panel. We then developed a scoring system that predicts an aptazymes's activity by integrating three characteristics of communication-module bases: hydrogen bonding, base stacking, and distance to the enzymatic core. We validated the power and generality of this approach by designing aptazymes responsive to three distinct ligands, each with markedly wider dynamic ranges than any previously reported. These aptayzmes efficiently regulated adeno-associated virus (AAV)-vectored transgene expression in cultured mammalian cells and mice, highlighting one application of these broadly usable regulatory switches. Our approach enables efficient, protein-independent control of gene expression by a range of small molecules.
Article and author information
Author details
Funding
National Institutes of Health (R01 AI091476)
- Michael Farzan
National Institutes of Health (P01 AI100263)
- Michael Farzan
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocol (#14-028) of the Scripps Florida.
Copyright
© 2016, Zhong et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Rational design of aptazyme riboswitches for efficient control of gene expression in mammalian cells
eLife 5:e18858.
https://doi.org/10.7554/eLife.18858
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