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March 1, 1997Journal of VirologyOpen Access

Analysis of recombinant adeno-associated virus packaging and requirements for rep and cap gene products

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Authors

KVKaren A. VincentSPSusan PirainoSWSamuel C. Wadsworth

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Overview

In vitro study demonstrates that elevating capsid protein synthesis boosts recombinant adeno-associated virus yield tenfold, indicating structural protein levels limit vector packaging.

Key Points

  • To determine whether the expression levels of replication (Rep) and capsid (Cap) proteins serve as limiting factors in recombinant adeno-associated virus (rAAV) packaging.
  • Engineered helper plasmids by replacing endogenous AAV p5 and p40 promoters with the Rous sarcoma virus LTR and cytomegalovirus immediate-early promoter, respectively.
  • Supplied Rep and Cap proteins in trans during adenovirus-assisted rAAV vector packaging to measure vector yield and transcriptional activity.
  • Increasing Cap protein synthesis via the cytomegalovirus promoter produced an approximately 10-fold increase in rAAV yield.
  • Expressing Rep proteins from the RSV LTR failed to increase rAAV yield and inhibited adenovirus-mediated activation of p40 transcription, ultimately lowering capsid protein synthesis.

Cite This Study

Vincent et al. (1997) studied this question.

synapsesocial.com/papers/6a70026635aa2c282ce1504ahttps://doi.org/10.1128/jvi.71.3.1897-1905.1997
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