Part:BBa_K209400

AarI B-C part, hM4D

hM4 DREADD flanked by AarI BC sites.

Usage and Biology

This is an engineered version of the human M4 muscarinic receptor that can be activated by the otherwise inert small molecule, clozapine-N-oxide.

Characterization by Team UCSF 2010

In 2009 we used this synthetic GPCR to mediate chemotaxis towards the ligand, CNO.
For more information, refer to our wiki here: http://2009.igem.org/Team:UCSF

In 2010, we showed that this synthetic GPCR increases killer cell activation in the presence of the ligand, CNO:

<img src="" />
Figure: Enhancing killer cell activation with a synthetic GPCR
A synthetic GPCR shown to mediate immune cell migration towards the ligand CNO was introduced to T cells as a potential way to increase the level of kinase activation. IL-2 production was used as a measure of T-cell activation. The results show that in the presence of CNO, T cells transfected with the synthetic GPCR have a higher level of activation than non-transfected T-cells.
For the complete results, refer to our wiki here: http://2010.igem.org/Team:UCSF/Project/Signaling

Reference: Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand. Armbruster BN, Li X, Pausch MH, Herlitze S, Roth BL. Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):5163-8. Epub 2007 Mar 2.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 184
  Illegal PstI site found at 979
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1215
  Illegal PstI site found at 184
  Illegal PstI site found at 979
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 184
  Illegal PstI site found at 979
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 184
  Illegal PstI site found at 979
  Illegal NgoMIV site found at 1086
  
• 1000
  COMPATIBLE WITH RFC[1000]