Nomeclature regarding AAV constructs

Q:  "On UNC vector core website (and DREADD wiki pages), I found two types, AAV2/5/8-CAMIIKa-HA-HM3Dq-IRES-mCitrine or AAV8-hsyn-HM3Dq-mCherry.

 My question is, which one is better? and can I know the difference between them, in the aspect of expression level or anatomical specificity?

 A:  The nomneclature gives the answer:

 AAV2/5/8-CAMIIka-HA-hM3Dq-IRES-mCitrene=

 This is available in three serotypes (AAV2, AAV5 and AAV8)=we recommend starting with AAV8 serotypes 
CAMKIIka=the promotor. In this case the promotor is selective for neurons which express CAMIIKa (e.g. glutamatergic neurons in general)

 HA=Hemagluttinin tag

 hM3Dq=Gq DREADD

 IRES=internal ribosome entry site

 mCitrene=GFP variant which is quite bright and more resistant to photobleaching than GFP
hSyn=human synapsin promotor which in this case specifies neuronal expression

 M3Dq-mCherry=Gq DREADD as mCherry fusion protein

 Thus, both will express Gq DREADD in neurons but one will specify excitatory neurons.

Comments

Post a Comment

Popular posts from this blog

CNO in drinking water

We frequently get requests regarding protocols for putting CNO in the drinking water for long-term studies. Here is our protocol: 1. Dissolve the CNO in a small volume of DMSO 2. Diluted the dissolved CNO in drinking water (e.g., 5mg/200ml) 3. Give mice with CNO drinking water  and protect from light using foil-wrapped bottles 4. The mice will receive 5mg/kg/day CNO (assume that mice weight 30g and consume 6ml water per day) 5. Prepare fresh daily. 6. A small amount of saccharine in the drinking water will mask the slightly bitter taste of CNO
Read more

How best to dissolve CNO ...it depends on what it is soluble in (and that depends on the particular polymorph)

I recently received the following question: " My question is with regard to the use of CNO in drinking water (in mouse studies). Your recipe calls for the addition of a small amount of DMSO to dissolve CNO at a concentration of 5 mg in 200 mL. As we and others routinely dissolve CNO in 0.9% saline (no DMSO) at concentrations up to four times this amount, I am curious whether the addition of DMSO to CNO solutions for H2O drinking is critical, or whether the properties of CNO permit it's direct dissolution in H2O at concentrations from 5-20 mg per 200 mL. There appears to be no difficulty dissolving such concentrations in H2O in my hands, but I would prefer to hear your perspective. Is there a published reference for the solubility profile of CNO in H2O or 0.9% saline?" It basically depends upon the source of the CNO.  CNO via the NIH RAID program (which is what we use) is not readily soluble in H20 while commercial CNO is.  Although we've verified that both source
Read more