Rational design of aptazyme riboswitches for efficient control of gene expression in mammalian cells
UMass Chan Affiliations
Gene Therapy CenterDocument Type
Journal ArticlePublication Date
2016-11-02Keywords
RNA switchaptazyme
biochemistry
gene regulation
human
mammalian cells
mouse
small molecule
Biochemistry
Genetics and Genomics
Metadata
Show full item recordAbstract
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.Source
Elife. 2016 Nov 2;5. pii: e18858. doi: 10.7554/eLife.18858. Link to article on publisher's siteDOI
10.7554/eLife.18858Permanent Link to this Item
http://hdl.handle.net/20.500.14038/40205PubMed ID
27805569Related Resources
Link to Article in PubMedRights
Copyright © 2016, Zhong et al.Distribution License
http://creativecommons.org/licenses/by/4.0/ae974a485f413a2113503eed53cd6c53
10.7554/eLife.18858