Team:William and Mary/parts

ChR1

Channelrhodopsin-1, or ChR1, is a light-gated ion channel. Responding to light of approximately 560 nm (yellow light), ChR1 is a non-specific cation channel, allowing the influx of ions such as calcium into cells. Unfortunately, ChR1 contained two internal PstI cutsites, which needed to be mutated out before it could be put into a biobrick backbone.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

See our part in the Registry at: parts.igem.org/Part:BBa_K1409000

This part is originally from the lab of Dr. Karl Deisseroth at Stanford University and was purchased from Addgene.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

ChR2

Channelrhodopsin-2 (ChR2), is another channelrhodopsin. The key differerence between ChR2 and ChR1 is that ChR2 absorbs blue light. ChR2 also contained an internal PstI cutsite that needed to be dealt with.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

This part is originally from the lab of Dr. Karel Svoboda at Janelia Farm and was purchased from Addgene.

CatCh

CatCh, or Calcium Relocating Channelrhodopsin, is a channelrhodopsin with an increased Ca2+ permeability. CatCh also had two internal PstI cutsites that were mutated out.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

See our part in the Registry at: parts.igem.org/Part:BBa_K1409001

This part is originally from the lab of Dr. Peter Hegemann at Humboldt-Universität in Berlin.

GCaMP/RCaMP

GCaMP, a Genetically Encoded Calcium Indicator (or GECI), is a protein created from the fusion of GFP, calmodulin (a calcium binding messenger protein), and M13 (a sequence from myosin light chain kinase). When calcium binds to CaM, conformational changes in the protein cause a change in the intensity of the fluorescence of GFP.

 

Functioning in much the same way as GCaMP, RCaMP is another GECI made from mRuby instead of GFP, and fluorescing red instead of green.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

The green fluorescence in this image is a result of the binding of calcium to GCAMP. This image was obtained from the lab of Dr. Margaret Saha at the College of William and Mary, from which we also acquired samples of GCaMP and RCaMP.