I am seeking to use in vivo injection of AAV in adult mice to express TEV protease in to the heart muscle cells of our mouse line. We do not have experience with this, but based on the literature, it seems others have done this successfully. However, researches don't seem to agree whether AAV6 or AAV9 works better. Have you done this before? what promoter did you use? Did you have high protein expression? Which serotype did you use? Any sort of advice would be greatly appreciated!! Thank you
Dear John Sullivan
Yes it is possible to successfully over-express the protein of interest using AAV particles.
If you wish to target the heart in an in vivo setting pls use AAV9 with a CMV (ubiquitous) promoter.
Further more if you aim to induce expression in a cell type specific manner then you can use other promoters which are specific for the target cell type of your choice. (For eg, to overexpress in cardiomyocytes you may use a cTNT promoter).
The time taken to achieve high over-expression will depend on the strength of the promoter used and also to a certain extent on the size of the protein being expressed.
However, pls consider the packaging limit of the AAV particles when you plan your experiments. If your transgene is larger than 3.5kb then it will not be possible to use AAV mediated over-expression. As long as your transgene of interest is within 3.5kb you can achieve AAV mediated over-expression.
Hope this was helpful.
Good luck !
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