We occasionally receive requests for the lentiviral vectors in Alexander et al, Neuron 2009. We will soon make them available via ADDGENE but for now simply contact Hu ( from the Rothlab
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
I receive frequent queries regarding the appropriate way to visualize DREADDs. Below I show the three basic DREADD constructs: HA-tagged mCherry fusion proteins HA-tagged in a vector with IRES-mCitrene In unfixed tissue the mCherry and mCitrene can be visualized using the parameters from the table below or by going here . NOTE: only the mCherry fusion protein fluorescence will show that the DREADD receptor is expressed. mCitrene simply will show which cells were transfected or transduced. For visualizing HA we have a detailed recommended protocol the principle of which is below. I also show various secondary antibodies which might be used along with excitation/emission characteristics Here's what you might consider if you have an mCherry fusion protein: Finally, what if you are using one of the HA-hM3Dq-IRES-mCitrene or HA-hM4Di-IRES-mCitrene constructs. ...
Although we've not yet published anything on these JAX would like us to let all know the following which lists the mice as well as direct links to the JAXMice datasheets. --JR#0262 19 R26-LSL-G i -DREADD (Rosa-CAG-LSL-HA-hM 4Di -pta- mCitrine) http://jaxmice.jax.org/strain/ 026219.html --JR#0262 20 R26-LSL-G q -DREADD (Rosa-CAG-LSL-HA-hM 3Dq -pta- mCitrine) http://jaxmice.jax.org/strain/ 026220.html --JR#0262 61 R26-LSL-G s -DREADD (Rosa-CAG-LSL-HA- r M 3Ds -pta- mCitrine) http://jaxmice.jax.org/strain/ 026261.html
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