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

Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Hungária krt. 21, H-1143 Budapest, Hungary
Hungarian National Blood Transfusion Service, Laboratory of Transplantation Immunogenetics, Karolina út 19-21, H-1113 Budapest, Hungary
Institute of Virology, Campus Benjamin Franklin, Charité Medical School, Hindenburgdamm 27, 12203 Berlin, Germany
Author to whom correspondence should be addressed.
Viruses 2019, 11(1), 38;
Received: 10 December 2018 / Revised: 4 January 2019 / Accepted: 4 January 2019 / Published: 9 January 2019
(This article belongs to the Special Issue New Insights into Parvovirus Research)
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. View Full-Text
Keywords: AAV2; adeno-associated virus; bisulfite PCR; CpG methylation; DNA virus; Parvoviridae AAV2; adeno-associated virus; bisulfite PCR; CpG methylation; DNA virus; Parvoviridae
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Tóth, R.; Mészáros, I.; Hüser, D.; Forró, B.; Marton, S.; Olasz, F.; Bányai, K.; Heilbronn, R.; Zádori, Z. Methylation Status of the Adeno-Associated Virus Type 2 (AAV2). Viruses 2019, 11, 38.

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