Synthetic long non-coding RNAs [SINEUPs] rescue defective gene expression in vivo.

Alessia Indrieri,Claudia Grimaldi,Roberta Tammaro,Stefano Gustincich,Brunella Franco,Silvia Zucchelli

doi:10.1038/srep27315

Abstract

Non-coding RNAs provide additional regulatory layers to gene expression as well as the potential to being exploited as therapeutic tools. Non-coding RNA-based therapeutic approaches have been attempted in dominant diseases, however their use for treatment of genetic diseases caused by insufficient gene dosage is currently more challenging. SINEUPs are long antisense non-coding RNAs that up-regulate translation in mammalian cells in a gene-specific manner, although, so far evidence of SINEUP efficacy has only been demonstrated in in vitro systems. We now show that synthetic SINEUPs effectively and specifically increase protein levels of a gene of interest in vivo. We demonstrated that SINEUPs rescue haploinsufficient gene dosage in a medakafish model of a human disorder leading to amelioration of the disease phenotype. Our results demonstrate that SINEUPs act through mechanisms conserved among vertebrates and that SINEUP technology can be successfully applied in vivo as a new research and therapeutic tool for gene-specific up-regulation of endogenous functional proteins.

Highlights

Both naturally occurring and artificial RNAs have the potential to be used as modulators of target genes
We demonstrated that AS Uchl[1] activity depends on two distinct RNA elements, The Binding Domain (BD) at the 5′ end, is a sequence that overlaps, in antisense orientation, to the sense protein-coding mRNA and determines target selection and AS Uchl[1] specificity by RNA-RNA base pairing
For our initial studies we utilized the SINEUP-GFP7 that results in an increase GFP protein levels in transient overexpression experiments

Summary

Introduction

Both naturally occurring and artificial RNAs have the potential to be used as modulators of target genes. We show for the first time that SINEUPs can increase the synthesis of a functional endogenous protein in vivo and rescue haploinsufficient gene dosage in a medakafish model of a human disease.

Results

Conclusion