Has anyone used EF1a promoters to drive transgene expression in DRGs in vivo? I’ve had success with hSyn, and recently, a paper from Haenraets et al showed CAG works well for DRG. But EF1a is also very popular and it’d be nice to use. I haven’t tried in vivo. EF1a works on cultured DRGs (I’ve tried this) but I’ve not seen any in vivo.
To answer my own question:
From Arcourt et al (2017):
They injected into the L3-L4 ganglia directly. I was thinking intrathecally. So that remains to be determined. But it looks like EF1a promoter is active in DRG.
Hi, Did you try the intrathecal injection with EF1a promoter? Did it work for you in DRG?
I tried. Didn’t look great but I don’t have much confidence in our viral prep in that instance. Try something from Addgene or UNC core and see how it looks. Let us know.
Hey! Did you ever give CAG or CBA a shot btw? We want to fit in DREADDs with a fluorophore but CAG is pretty large so we might go with CBA. I also recently saw this paper — it’s from a former member of Feng Zhang’s lab. https://www.sciencedirect.com/science/article/pii/S2211124723013608?via%3Dihub
Supposedly there are some new AAV serotypes (MaCPNS1 and MaCPNS2) from the Gradinaru lab which have better transduction results in DRG/Nodose – we’re interested in testing one of those as well. https://www.sciencedirect.com/science/article/pii/S0896627322004111
CAG is probably most dependable. BUt if you want to consider CBA, get an off the shelf vector with CBA, try it, see how it expresses.
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Sweet, we ended up just yolo-ing and put in an order from VectorBuilder for a cre-dependent hm4di-mScarlet3 construct under CBA promoter, with AAV9 or MaCPNS1 serotype. I’ll let yall know how it goes. CAG would have been ideal but we wanted to fit the fluorphore in. If CBA doesnt work and we try CAG, I suppose we could swap the fluorophore out for a stainable his-tag or something similar (though I’m not sure how well IHC staining usually works for these – I would prefer not to deal with amplification kits)