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.
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.
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