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Welcome to the Designer Receptors Exclusively Activated by Designer Drugs DREADD wiki

For the most recent updates, available resources and protocols on DREADD technology please visit our blog http://chemogenetic.blogspot.com/

DREADDs are engineered G-protein coupled receptors which are activated by otherwise inert drug-like small molecules. In the example shown below, an evolved M3-muscarinic receptor is activated by the small molecule clozapine-N-oxide (CNO; see Armbruster et al, PNAS 2007 for details) http://www.ncbi.nlm.nih.gov/pubmed/17360345

http://pharmrev.aspetjournals.org/content/63/2/291/F3.small.gif

DREADDs are currently used by many labs to remotely and non-invasively control neuronal signaling

http://davecharest.com/wp-content/uploads/2009/12/neurons1-240x180.jpg

Helpful reviews on the technology:

Rogan and Roth, Pharmacol Rev 2011 Jun;63(2):291-315. Epub 2011 Mar 17. http://www.ncbi.nlm.nih.gov/pubmed/21415127

Dong et al, Mol Biosystems 2010 Aug;6(8):1376-80. Epub 2010 Jun 7. http://www.ncbi.nlm.nih.gov/pubmed/20532295


Nomenclature

Gq-DREADD = hM3Dq = D-q ; clozapine-N-oxide activates Gq-mediated signaling. In neurons activation of hM3Dq induces burst firing

Gi-DREADD=hM4Di= D-i; clozapine-N-oxide induces Gi activation. In neurons, activation of hM4Di induces neuronal silencing

Gs-DREADD = Gs-D = D-s; clozapine-N-oxide induces Gs activation. In neurons this leads to an increase in cAMP.

Detailed Protocols for Creating DREADDs

Dong et al, Nature Protocols 2010;5(3):561-73. Epub 2010 Feb 25. http://www.ncbi.nlm.nih.gov/pubmed/20203671


Listed below are various DREADD constructs currently available with links to VectorNTI files:

available in pcDNA3.0:

HA-tagged hM3D in pcDNA3: this is coupled to Gq and induces neuronal burst firinghttp://www.ncbi.nlm.nih.gov/pubmed/19607790

HA-tagged hM4D in pcDNA3: this is coupled to Gi and induces neuronal silencing http://www.ncbi.nlm.nih.gov/pubmed/17360345

HA-tagged Gs-DREADD in pcDNA3: this is coupled to Gs and elevates cAMP production http://www.ncbi.nlm.nih.gov/pubmed/19858481



Available as FLEX-AAV Constructs using Synapsin-1 promotor (also known as DIO Vectors) (See Krashes et al, JCI 2011) http://www.ncbi.nlm.nih.gov/pubmed/21364278)
mCherry Fusion Proteins

hM3Dq-mCherry fusion protein construct http://www.ncbi.nlm.nih.gov/pubmed/21364278: Causes burst firing of neurons

hM4Di-mCherry fusion protein construct http://www.ncbi.nlm.nih.gov/pubmed/21364278: Silences neuronal firing


AAV-FLEX Constructs with Syanpsin-1 promotor; also contain IRES and either mCherry or mCitrine reporters: Validated in HEK-293 cells and neurons but not yet published.

HIGH TITER VIRUS NOW AVAILABLE FROM THE UNC VECTOR CORE FOR mCITRENE VERSIONS
HA-hM3Dq-IRES-mCherry

HA-hM3Dq-IRES-mCitrine


HA-hM4Di-IRES-mCherry

HA-hM4Di-IRES-mCitrine



How to obtain high-titer virus stocks: Each aliquot is ~$220 and no MTA needed. How to obtain:

  1. Please go to http://genetherapy.unc.edu/services.htm
  1. Fill out the order form for serotype 5, 2 or 8 virus. We find that AAV serotype 8 yields highest levels of neuronal expression and greatest spread of infection and is ideal for studies using DIO vectors. Serotypes 2 and 5 are best where localized expression is key.

Available in Lentiviral vectors

HA-hM3Dq-IRES-mCherry http://www.ncbi.nlm.nih.gov/pubmed/19607790

HA-hM4Dq-IRES-mCherry

DREADDs targeted to neuronal subdomains (available for collaborative studies only):

Somato-dendritically-targeted DREADDs (hM4Di, hM3Dq)

Axonally-targeted DREADD (hM4Di, hM3Dq)

Glial-specific expression: AAV-GFAP minimal promotor available via UNC Vector core

Glial-specific Gs and Gq DREADDs (GsD and hM3Dq)


Mice currently available:

tet-HA-tagged hM3Dq mice: http://www.ncbi.nlm.nih.gov/pubmed/19607790
tet-HA-tagged hM4Dq mice: http://www.ncbi.nlm.nih.gov/pubmed/24525710

Adora2A-GsD mice:http://www.ncbi.nlm.nih.gov/pubmed/23303063

http://www.antibodyreview.com/article_images/19893765/Front%20Mol%20Neurosci/2-_/p16-2773177/fnmol-02-016-g002.jpg



The Jackson lab offers tet- HA-tagged hM4Di mice http://jaxmice.jax.org/strain/024114.html

The Jackson Laboratory offers tet-HA-tagged hM3Dq mice: http://jaxmice.jax.org/strain/014093.html

Adora2A-rM3Ds-mCherry mice: http://jaxmice.jax.org/strain/017863.html

Detailed protocol for HA-staining now available (see attachments below)

Detailed protocol for dissolving CNO now available (see attachments below)

Vector maps available for common vectors (see Attachments)

Last modified 16 months ago Last modified on 02/06/15 13:49:59

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