Transgene Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors

Gene therapy uses a viral capsid to introduce DNA into cells, and then the cells process that DNA through mechanisms that aren't fully understood. By studying how the cell processes AAV delivered DNA, we hope to improve safety, efficacy and longevity of AAV gene therapies. Studying AAV DNA processing at the tissue or organism level was quite difficult.

And while we were developing cell culture transduction protocols for AAV visualization via super resolution microscopy, we found that the simplicity of producing and using unpurified AAV preparations in cell culture is not well known. This straightforward method for transient expression can be advantageous for researchers in numerous fields. Creating crude AAV preparations is quite simple, and they have a long shelf life F4C, allowing them to be used in numerous experiments.

Applying them is simple, making them a time and cost-effective alternative to standard transfection reagents. Often AAV crude preparations are more efficient and less toxic than transfection. Vectrology has two arms, production and transduction, and both are entirely dependent on host cellular machinery.

Our future research aims to identify the cellular machinery involved in these processes and ways to manipulate them so that we can improve production and transduction efficiency. Understanding the basic biology underpinning both arms of vectrology is vital for the success of gene therapies.