Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated viral vectors and improved alternative methods.

@article{Fagone2012SystemicEI,
  title={Systemic errors in quantitative polymerase chain reaction titration of self-complementary adeno-associated viral vectors and improved alternative methods.},
  author={Paolo Fagone and J. F. Wright and Amit Chunilal Nathwani and A. W. Nienhuis and Andrew M. Davidoff and John Mark Nicholas Timm Gray},
  journal={Human gene therapy methods},
  year={2012},
  volume={23 1},
  pages={1-7}
}
Self-complementary AAV (scAAV) vector genomes contain a covalently closed hairpin derived from a mutated inverted terminal repeat that connects the two monomer single-stranded genomes into a head-to-head or tail-to-tail dimer. We found that during quantitative PCR (qPCR) this structure inhibits the amplification of proximal amplicons and causes the systemic underreporting of copy number by as much as 10-fold. We show that cleavage of scAAV vector genomes with restriction endonuclease to… CONTINUE READING
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Early clinical trial results following administration of a low dose of a novel self complementary adeno associated viral vector

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