Generation of high-quality single-stranded DNA for full-length and truncated genome standards of recombinant adeno-associated viruses.

ABSTRACT: The integrity of the single-stranded DNA (ssDNA) genome of recombinant adeno-associated viral vectors (rAAVs) for gene therapy applications is a critical quality attribute. To monitor this attribute, robust analytical methods, such as long-read sequencing or multiplex PCR, are needed. Due to the heterogeneity of the packaged rAAV payloads, the availability of appropriate ssDNA standards could clearly facilitate the development of these methods and may be used for system suitability testing. However, due to a genome length of up to approximately 4.7 kilobases and the secondary structure of the inverted terminal repeats, such standards are not trivial to produce. Here, we introduce a versatile method that starts with plasmid DNA containing the rAAV genome of interest. After its linearization

SUBMITTER: Manz J 

PROVIDER: S-EPMC12552963 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature