DataVXACS NanoPaper
journal article·2023-12-12

Engineering U1-Based Tetracycline-Inducible Riboswitches to Control Gene Expression in Mammals

Eric Rovira, Beatriz Moreno, Nerea Razquin, Lorea Blázquez, Ruben Hernández-Alcoceba, Puri Fortes, Fernando Pastor

ACS Nano· Volume 17· Issue 23
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Source abstract

Abstract

Synthetic riboswitches are promising regulatory devices due to their small size, lack of immunogenicity, and ability to fine-tune gene expression in the absence of exogenous trans-acting factors. Based on a gene inhibitory system developed at our lab, termed U1snRNP interference (U1i), we developed tetracycline (TC)-inducible riboswitches that modulate mRNA polyadenylation through selective U1 snRNP recruitment. First, we engineered different TC-U1i riboswitches, which repress gene expression unless TC is added, leading to inductions of gene expression of 3-to-4-fold. Second, we developed a technique called Systematic Evolution of Riboswitches by Exponential Enrichment (SEREX), to isolate riboswitches with enhanced U1 snRNP binding capacity and activity, achieving inducibilities of up to 8-fold. Interestingly, by multiplexing riboswitches we increased inductions up to 37-fold. Finally, we demonstrated that U1i-based riboswitches are dose-dependent and reversible and can regulate the expression of reporter and endogenous genes in culture cells and mouse models, resulting in attractive systems for gene therapy applications. Our work probes SEREX as a much-needed technology for the in vitro identification of riboswitches capable of regulating gene expression in vivo .
Source: openalex_inverted_index · Quality: baseline

Research context

OpenAlex topics: RNA and protein synthesis mechanisms · RNA Research and Splicing · RNA Interference and Gene Delivery

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Gene Therapy and Regulation of Gene Expression, Center for Applied Medical Research (CIMA), University of Navarra (UNAV), Pamplona 31008, Spain

Department of Molecular Therapy, Aptamer Unit, Center for Applied Medical Research (CIMA), University of Navarra (UNAV), Pamplona 31008, Spain

Department of Neurosciences, Biodonostia Health Research Institute, 20014 San Sebastián, Spain

CIBERNED, ISCIII (CIBER, Carlos III Institute, Spanish Ministry of Sciences and Innovation), 28031 Madrid, Spain

Ikerbasque, Basque Foundation for Science, 48009 Bilbao, Spain

Navarra Institute for Health Research (IdiSNA), Pamplona 31008, Spain

Spanish Network for Advanced Therapies (TERAV ISCIII), Madrid 28029, Spain

Liver and Digestive Diseases Networking Biomedical Research Centre (CIBERehd), Madrid 28029, Spain

Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid 28029, Spain

Funding

FP7 People: Marie-Curie Actions · European Commission · Ministerio de Ciencia e Innovaci?n · European Regional Development Fund · Ikerbasque, Basque Foundation for Science · Fundaci?n Cient?fica Asociaci?n Espa?ola Contra el C?ncer · 'la Caixa' Foundation · Gobierno de Navarra · Instituto de Salud Carlos III

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