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

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.

miR‐181a/b downregulation exerts a protective action on mitochondrial disease models

“…We previously generated two medakafish ( Oryzias latipes ) models of MLS by knocking down, using a Morpholino(MO)‐based approach, hccs or cox7B expression (Indrieri et al , , ). Both models ( hccs ‐MO and cox7B ‐MO) showed a severe microphthalmic and microcephalic phenotype due to increased cell death in the CNS (Indrieri et al , , , ). In medaka, the mature forms of miR‐181a and miR‐181b are perfectly conserved with respect to their mammalian counterparts, in terms of both sequence identity (100%) and pattern of expression in the retina and brain (Carrella et al , ).…”

The Pervasive Role of the miR-181 Family in Development, Neurodegeneration, and Cancer

“…The authors showed that miR-181a and b control global regulation of mitochondrial turnover in the CNS through coordinated activation of mitochondrial biogenesis and mitophagy [28]. They demonstrated that the silencing of these two miRNAs significantly rescues neurodegeneration in two fish models of Microphthalmia with linear skin defects syndrome (MLS; MIM309801 [136]), a rare neurodevelopmental disorder due to mutations in players of the MRC Complex III and IV [137][138][139]. Moreover, they tested the effects of genetic inactivation of miR-181a and b in a chemical (rotenone-induced [140]) and genetic (Ndufs4 -/- [141]) murine models of Leber hereditary optic neuropathy (LHON, MIM535000 [142]), a mitochondrial disorder characterized by degeneration of RGCs and loss of central vision.…”

Engineering Translation in Mammalian Cell Factories to Increase Protein Yield: The Unexpected Use of Long Non-Coding SINEUP RNAs

“…In our recent work, we designed synthetic SINEUPs to target endogenous DJ-1 mRNA, a gene involved in recessive familial forms of Parkinson's Disease, and we could knock-up endogenous DJ-1 protein levels up to 3-fold in 3 different neuronal cell lines in vitro [97]. Subsequently, in a collaborative effort aimed at proving that SINEUP technology can also be applied in vivo , we could rescue the defective gene expression in a medakafish model of Microphtalmia with Linear Skin Lesion, a human disorder characterized by haploinsufficient dosage of COX7b protein [99].…”