Infectivity of rAAV vectors is a key attribute impacting the efficacy of AAV mediated gene therapy, however, it has been challenging to determine rAAV infectious titres as titration methods generally rely on the detection of a reporter marker or amplification of the rAAV genome for reliable detection within an infected cell. Use of wildtype AAVs or specific engineered cell lines (e.g. HeLaRC32) with wildtype adenovirus serotype 5 (Ad5) to supply the necessary components to replicate the rAAV genome are commonly employed for infectious titration of rAAV. However, these methods raise safety concerns associated with the use of wildtype viruses and how representative the titres are in the target cell types. Here we developed a novel rAAV infectious assay based on our self-silencing adenovirus platform entitled ‘Tetracycline-Enabled Self-Silencing Adenovirus’ (TESSA).
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