Can I validate target gene expression by transfecting pAAV-hSyn-TALE-VP64/SIDX4-2A-EGFP constructs into Neuro2A cells?

[live2l...@gmail.com]

Dear Forum members,

I am planning to transfect these pAAV-hSyn-TALE-VP64/SIDX4-2A-EGFP constructs into N2A cells to check if they work prior to AAV production. Has anyone done this in N2A cells before? My concern is that the hSyn may not work well in N2As. If anyone has done this, can you see EGFP expression in live cells?

Thanks for your help!

-Garrett

[Le Cong]

Hi Garrett,

The hSyn promoter should have some activity in N2A cells. I have tested similar constructs and saw GFP signal, though it's relatively weak. But because the construct I used is not exactly the same you might get different results. I do think you could give it a try while keep in mind the signal would be weaker than if you make AAV.

Best,

Le

Le CONG, Ph.D. 丛乐

Broad Institute of MIT and Harvard,

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[Garrett Kaas]

So, just to follow up on the use of N2A cells for pAAV-hSyn-TALE-VP64/SIDX4 2A-EGFP construct validation. I tried it and it worked great--GFP signal was clearly visible in live cells. Thanks for your help. 

Garrett

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