Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle.
Recombinant adeno-associated viral (rAAV) vectors can support long-term transgene expression in quiescent tissues. Intramuscular (i.m.) administration of a single-stranded AAV vector (ssAAV) in the nonhuman primate (NHP) results in a peak protein level at 2-3 months, followed by a decrease over seve...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3110818?pdf=render |
| _version_ | 1818542026370580480 |
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| author | Adrien Léger Caroline Le Guiner Michael L Nickerson Kate McGee Im Nicolas Ferry Philippe Moullier Richard O Snyder Magalie Penaud-Budloo |
| author_facet | Adrien Léger Caroline Le Guiner Michael L Nickerson Kate McGee Im Nicolas Ferry Philippe Moullier Richard O Snyder Magalie Penaud-Budloo |
| author_sort | Adrien Léger |
| collection | DOAJ |
| description | Recombinant adeno-associated viral (rAAV) vectors can support long-term transgene expression in quiescent tissues. Intramuscular (i.m.) administration of a single-stranded AAV vector (ssAAV) in the nonhuman primate (NHP) results in a peak protein level at 2-3 months, followed by a decrease over several months before reaching a steady-state. To investigate transgene expression and vector genome persistence, we previously demonstrated that rAAV vector genomes associate with histones and form a chromatin structure in NHP skeletal muscle more than one year after injection. In the mammalian nucleus, chromatin remodeling via epigenetic modifications plays key role in transcriptional regulation. Among those, CpG hyper-methylation of promoters is a known hallmark of gene silencing. To assess the involvement of DNA methylation on the transgene expression, we injected NHP via the i.m. or the intravenous (i.v.) route with a recombinant ssAAV2/1 vector. The expression cassette contains the transgene under the transcriptional control of the constitutive Rous Sarcoma Virus promoter (RSVp). Total DNA isolated from NHP muscle and liver biopsies from 1 to 37 months post-injection was treated with sodium bisulfite and subsequently analyzed by pyrosequencing. No significant CpG methylation of the RSVp was found in rAAV virions or in vector DNA isolated from NHP transduced tissues. Direct de novo DNA methylation appears not to be involved in repressing transgene expression in NHP after gene transfer mediated by ssAAV vectors. The study presented here examines host/vector interactions and the impact on transgene expression in a clinically relevant model. |
| first_indexed | 2024-12-11T22:16:46Z |
| format | Article |
| id | doaj.art-dce465869f324f5f90b1718ab716eb4b |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T22:16:46Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-dce465869f324f5f90b1718ab716eb4b2022-12-22T00:48:35ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0166e2088110.1371/journal.pone.0020881Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle.Adrien LégerCaroline Le GuinerMichael L NickersonKate McGee ImNicolas FerryPhilippe MoullierRichard O SnyderMagalie Penaud-BudlooRecombinant adeno-associated viral (rAAV) vectors can support long-term transgene expression in quiescent tissues. Intramuscular (i.m.) administration of a single-stranded AAV vector (ssAAV) in the nonhuman primate (NHP) results in a peak protein level at 2-3 months, followed by a decrease over several months before reaching a steady-state. To investigate transgene expression and vector genome persistence, we previously demonstrated that rAAV vector genomes associate with histones and form a chromatin structure in NHP skeletal muscle more than one year after injection. In the mammalian nucleus, chromatin remodeling via epigenetic modifications plays key role in transcriptional regulation. Among those, CpG hyper-methylation of promoters is a known hallmark of gene silencing. To assess the involvement of DNA methylation on the transgene expression, we injected NHP via the i.m. or the intravenous (i.v.) route with a recombinant ssAAV2/1 vector. The expression cassette contains the transgene under the transcriptional control of the constitutive Rous Sarcoma Virus promoter (RSVp). Total DNA isolated from NHP muscle and liver biopsies from 1 to 37 months post-injection was treated with sodium bisulfite and subsequently analyzed by pyrosequencing. No significant CpG methylation of the RSVp was found in rAAV virions or in vector DNA isolated from NHP transduced tissues. Direct de novo DNA methylation appears not to be involved in repressing transgene expression in NHP after gene transfer mediated by ssAAV vectors. The study presented here examines host/vector interactions and the impact on transgene expression in a clinically relevant model.http://europepmc.org/articles/PMC3110818?pdf=render |
| spellingShingle | Adrien Léger Caroline Le Guiner Michael L Nickerson Kate McGee Im Nicolas Ferry Philippe Moullier Richard O Snyder Magalie Penaud-Budloo Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle. PLoS ONE |
| title | Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle. |
| title_full | Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle. |
| title_fullStr | Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle. |
| title_full_unstemmed | Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle. |
| title_short | Adeno-associated viral vector-mediated transgene expression is independent of DNA methylation in primate liver and skeletal muscle. |
| title_sort | adeno associated viral vector mediated transgene expression is independent of dna methylation in primate liver and skeletal muscle |
| url | http://europepmc.org/articles/PMC3110818?pdf=render |
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