Viral vectors, including adeno-associated vector (AAV), are an effective tool to deliver transgenes into the host cells. The frequency of vector transduction of targeted cells and organs can affect the efficacy, and there are two key ways to measure success: vector copy number (VCN) and transgene expression. VCN can be determined by extracting genomic DNA (gDNA) from bulk cells/tissue and utilizing quantitative PCR (qPCR) to determine the total number of viral genomes.
In the preclinical study phase, VCN and transgene expression are often tested from the same sample to ensure the accuracy and reproducibility of treatment performance. To do so, both DNA and RNA are extracted from the same biologic sample. Some existing protocols achieve simultaneous DNA and RNA extraction by splitting the lysate into two portions, then treating the portions of lysate with either DNase or RNase A. The drawback of this method, however, especially for precious samples, is that half of the DNA or RNA is lost.
This application note from Beckman Coulter Life Sciences demonstrates a simultaneous high-quality DNA and RNA extraction method for vector copy number and transgene expression testing that does not require splitting the sample lysate and shows consistent performance across species with 10 different tissue types and nine regions of the brain.