DREADD + CRISPR + iPS cells: cool paper

An interesting paper and another proof-of-concept study combining chemogenetics, CRISPR and iPS cell technology.

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

Considerations for immunofluorescence for various DREADD constructs

Image
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. ...
Read more

Floxed DREADD mice now available from JAX

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  
Read more