Domenger, C;
Allais, M;
François, V;
Léger, A;
Lecomte, E;
Montus, M;
Servais, L;
... Le Guiner, C; + view all
(2018)
RNA-Seq Analysis of an Antisense Sequence Optimized for Exon Skipping in Duchenne Patients Reveals No Off-Target Effect.
Molecular Therapy: Nucleic Acids
, 10
pp. 277-291.
10.1016/j.omtn.2017.12.008.
Preview |
Text
1-s2.0-S2162253117303074-main.pdf - Published Version Download (2MB) | Preview |
Abstract
Non-coding uridine-rich small nuclear RNAs (UsnRNAs) have emerged in recent years as effective tools for exon skipping for the treatment of Duchenne muscular dystrophy (DMD), a degenerative muscular genetic disorder. We recently showed the high capacity of a recombinant adeno-associated virus (rAAV)-U7snRNA vector to restore the reading frame of the DMD mRNA in the muscles of DMD dogs. We are now moving toward a phase I/II clinical trial with an rAAV-U7snRNA-E53, carrying an antisense sequence designed to hybridize exon 53 of the human DMD messenger. As observed for genome-editing tools, antisense sequences present a risk of off-target effects, reflecting partial hybridization onto unintended transcripts. To characterize the clinical antisense sequence, we studied its expression and explored the occurrence of its off-target effects in human in vitro models of skeletal muscle and liver. We presented a comprehensive methodology combining RNA sequencing and in silico filtering to analyze off-targets. We showed that U7snRNA-E53 induced the effective exon skipping of the DMD transcript without inducing the notable deregulation of transcripts in human cells, neither at gene expression nor at the mRNA splicing level. Altogether, these results suggest that the use of the rAAV-U7snRNA-E53 vector for exon skipping could be safe in eligible DMD patients.
| Type: | Article |
|---|---|
| Title: | RNA-Seq Analysis of an Antisense Sequence Optimized for Exon Skipping in Duchenne Patients Reveals No Off-Target Effect |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.omtn.2017.12.008 |
| Publisher version: | https://doi.org/10.1016/j.omtn.2017.12.008 |
| Language: | English |
| Additional information: | Copyright © 2017 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
| Keywords: | exon skipping, off-target, RNA sequencing, Duchenne, U7snRNA |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Developmental Neurosciences Dept |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10089547 |
Loading...
Loading...Loading...| 1. |  United States | 4 |
| 2. |  China | 2 |
| 3. |  United Kingdom | 1 |
Archive Staff Only
 |
View Item |
