Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation
Manipulation of pre-mRNA processing is a promising approach toward overcoming disease-causing mutations and treating human diseases. We show that a combined treatment applying two splice-manipulating technologies improves therapeutic efficacies to correct mutation-induced splice defects. Previously,...
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doaj-74d2e42aad1243f58059fb9674fc0ac22020-11-24T21:23:15ZengElsevierMolecular Therapy: Nucleic Acids2162-25312019-12-0118123130Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site MutationSaskia Breuel0Mariann Vorm1Anja U. Bräuer2Marta Owczarek-Lipska3John Neidhardt4Human Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, GermanyHuman Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, GermanyAnatomy, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, Germany; Research Center Neurosensory Science, University of Oldenburg, GermanyHuman Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, GermanyHuman Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, Germany; Research Center Neurosensory Science, University of Oldenburg, Germany; Joint research training group of the Faculty of Medicine and Health Sciences, University of Oldenburg, Germany and the University Medical Center Groningen, Groningen, Netherlands; Corresponding author: Prof. Dr. John Neidhardt, PhD, Human Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Ammerländer Heerstrasse 114-118, 26129 Oldenburg, Germany.Manipulation of pre-mRNA processing is a promising approach toward overcoming disease-causing mutations and treating human diseases. We show that a combined treatment applying two splice-manipulating technologies improves therapeutic efficacies to correct mutation-induced splice defects. Previously, we identified a family affected by retinitis pigmentosa caused by the homozygous BBS1 splice donor site mutation c.479G > A. The mutation leads to both exon 5 skipping and intron 5 retention. We developed a therapeutic approach applying lentivirus-mediated gene delivery of engineered U1 small nuclear RNA (U1), which resulted in increased levels of correctly spliced BBS1. Herein, we show that the therapeutic effect of the engineered U1 efficiently reverted exon skipping but failed to reduce the intron retention. To complement the engineered U1 treatment, we identified four different antisense oligonucleotides (AONs) that block intron 5 retention in BBS1 transcripts. A treatment using engineered U1 in combination with AONs showed the highest therapeutic efficacy and increased the amount of correctly spliced BBS1 transcripts. We did not detect elevated levels of apoptotic cell death in AON-treated cell lines. In conclusion, engineered U1 or AONs provide efficient therapies with complementary effects and can be combined to increase efficacy of therapeutic approaches to correct splice defects. Keywords: splicing, splice defect, AON, antisense oligonucleotide, U1 snRNA, gene therapy, BBS1, mutation, genetic therapy, Bardet-Biedl Syndrome 1http://www.sciencedirect.com/science/article/pii/S2162253119302276 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Saskia Breuel Mariann Vorm Anja U. Bräuer Marta Owczarek-Lipska John Neidhardt |
spellingShingle |
Saskia Breuel Mariann Vorm Anja U. Bräuer Marta Owczarek-Lipska John Neidhardt Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation Molecular Therapy: Nucleic Acids |
author_facet |
Saskia Breuel Mariann Vorm Anja U. Bräuer Marta Owczarek-Lipska John Neidhardt |
author_sort |
Saskia Breuel |
title |
Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation |
title_short |
Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation |
title_full |
Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation |
title_fullStr |
Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation |
title_full_unstemmed |
Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation |
title_sort |
combining engineered u1 snrna and antisense oligonucleotides to improve the treatment of a bbs1 splice site mutation |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2019-12-01 |
description |
Manipulation of pre-mRNA processing is a promising approach toward overcoming disease-causing mutations and treating human diseases. We show that a combined treatment applying two splice-manipulating technologies improves therapeutic efficacies to correct mutation-induced splice defects. Previously, we identified a family affected by retinitis pigmentosa caused by the homozygous BBS1 splice donor site mutation c.479G > A. The mutation leads to both exon 5 skipping and intron 5 retention. We developed a therapeutic approach applying lentivirus-mediated gene delivery of engineered U1 small nuclear RNA (U1), which resulted in increased levels of correctly spliced BBS1. Herein, we show that the therapeutic effect of the engineered U1 efficiently reverted exon skipping but failed to reduce the intron retention. To complement the engineered U1 treatment, we identified four different antisense oligonucleotides (AONs) that block intron 5 retention in BBS1 transcripts. A treatment using engineered U1 in combination with AONs showed the highest therapeutic efficacy and increased the amount of correctly spliced BBS1 transcripts. We did not detect elevated levels of apoptotic cell death in AON-treated cell lines. In conclusion, engineered U1 or AONs provide efficient therapies with complementary effects and can be combined to increase efficacy of therapeutic approaches to correct splice defects. Keywords: splicing, splice defect, AON, antisense oligonucleotide, U1 snRNA, gene therapy, BBS1, mutation, genetic therapy, Bardet-Biedl Syndrome 1 |
url |
http://www.sciencedirect.com/science/article/pii/S2162253119302276 |
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