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Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)

To analyze the methylation status of wild-type adeno-associated virus type 2 (AAV2), bisulfite PCR sequencing (BPS) of the packaged viral genome and its integrated form was performed and 262 of the total 266 CG dinucleotides (CpG) were mapped. In virion-packaged DNA, the ratio of the methylated cyto...

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Autores principales: Tóth, Renáta, Mészáros, István, Hüser, Daniela, Forró, Barbara, Marton, Szilvia, Olasz, Ferenc, Bányai, Krisztián, Heilbronn, Regine, Zádori, Zoltán
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6356613/
https://www.ncbi.nlm.nih.gov/pubmed/30634383
http://dx.doi.org/10.3390/v11010038
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author Tóth, Renáta
Mészáros, István
Hüser, Daniela
Forró, Barbara
Marton, Szilvia
Olasz, Ferenc
Bányai, Krisztián
Heilbronn, Regine
Zádori, Zoltán
author_facet Tóth, Renáta
Mészáros, István
Hüser, Daniela
Forró, Barbara
Marton, Szilvia
Olasz, Ferenc
Bányai, Krisztián
Heilbronn, Regine
Zádori, Zoltán
author_sort Tóth, Renáta
collection PubMed
description To analyze the methylation status of wild-type adeno-associated virus type 2 (AAV2), bisulfite PCR sequencing (BPS) of the packaged viral genome and its integrated form was performed and 262 of the total 266 CG dinucleotides (CpG) were mapped. In virion-packaged DNA, the ratio of the methylated cytosines ranged between 0–1.7%. In contrast, the chromosomally integrated AAV2 genome was hypermethylated with an average of 76% methylation per CpG site. The methylation level showed local minimums around the four known AAV2 promoters. To study the effect of methylation on viral rescue and replication, the replication initiation capability of CpG methylated and non-CpG methylated AAV DNA was compared. The in vitro hypermethylation of the viral genome does not inhibit its rescue and replication from a plasmid transfected into cells. This insensitivity of the viral replicative machinery to methylation may permit the rescue of the integrated heavily methylated AAV genome from the host’s chromosomes.
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spelling pubmed-63566132019-02-05 Methylation Status of the Adeno-Associated Virus Type 2 (AAV2) Tóth, Renáta Mészáros, István Hüser, Daniela Forró, Barbara Marton, Szilvia Olasz, Ferenc Bányai, Krisztián Heilbronn, Regine Zádori, Zoltán Viruses Communication To analyze the methylation status of wild-type adeno-associated virus type 2 (AAV2), bisulfite PCR sequencing (BPS) of the packaged viral genome and its integrated form was performed and 262 of the total 266 CG dinucleotides (CpG) were mapped. In virion-packaged DNA, the ratio of the methylated cytosines ranged between 0–1.7%. In contrast, the chromosomally integrated AAV2 genome was hypermethylated with an average of 76% methylation per CpG site. The methylation level showed local minimums around the four known AAV2 promoters. To study the effect of methylation on viral rescue and replication, the replication initiation capability of CpG methylated and non-CpG methylated AAV DNA was compared. The in vitro hypermethylation of the viral genome does not inhibit its rescue and replication from a plasmid transfected into cells. This insensitivity of the viral replicative machinery to methylation may permit the rescue of the integrated heavily methylated AAV genome from the host’s chromosomes. MDPI 2019-01-09 /pmc/articles/PMC6356613/ /pubmed/30634383 http://dx.doi.org/10.3390/v11010038 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Tóth, Renáta
Mészáros, István
Hüser, Daniela
Forró, Barbara
Marton, Szilvia
Olasz, Ferenc
Bányai, Krisztián
Heilbronn, Regine
Zádori, Zoltán
Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)
title Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)
title_full Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)
title_fullStr Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)
title_full_unstemmed Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)
title_short Methylation Status of the Adeno-Associated Virus Type 2 (AAV2)
title_sort methylation status of the adeno-associated virus type 2 (aav2)
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6356613/
https://www.ncbi.nlm.nih.gov/pubmed/30634383
http://dx.doi.org/10.3390/v11010038
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