Difference between revisions of "Part:BBa K2492001"

Revision as of 22:56, 1 November 2017

CoCHR Channelrhodopsin

The central part of our project. Part BBa_K2492001 contains CoCHR(Chloromonas oogama channelrhodopsin) [1]. In our project, CoCHR is a cation-selective light sensor, which promptly generates a large permeability of divalent cations after absorbing photons from our excitation light source. In our project, we hope to investigate how olfactory messages are transmitted by the neuron systems. In order to decrease delay or response comparing to chemical stimulus, we choose optogenetic techniques. Since C.elegans do not have sense to light, we express channelrodopsins in pairs of neuron as light receptors. CoCHR is the channelrodopsin in transgenic AWA. In order to express CoCHR in AWA specifically, we express CoCHR through the linkage to the AWA cell-specific promoter odr-10. Thus, when we shed ~470nm excitation light to transgenic C.elegans, the activated AWA is able to cause obvious response such as body swimming towards. While wild-type C.elegans not behave abnormally under the same stimulation. Whether the certain neuron is activated or not is indicated by GEM-GECO, a calcium indicator. In short, we established a light-sensed AWA controlling system, so as to utilize this part to manipulate the activation of AWA, establish the response to input stimulation. The merit of CoCHR is its sensitivity to light which is five-times than formal ChR2. Thus, the low intensity requirement prevents the cross-activation of Crimson(Chlamydomonas noctigama channelrhodopsin). Chrimson is another integrated channelrhodopsin in AWB. (Moreover, in order to indicate where the target neurons are. We also construct a fusion reporter protein called mCherry. At the same time, the fusion protein is allowed to estimate the expression level of CoCHR.)

[1]Schild, L. C., & Glauser, D. A. (2015). Dual color neural activation and behavior control with chrimson and cochr in caenorhabditis elegans. Genetics, 200(4), 1029-34.

[2] Nagel G, Ollig D, Fuhrmann M, Kateriya S, Musti AM, Bamberg E, Hegemann P (June 2002). "Channelrhodopsin-1: a light-gated proton channel in green algae". Science 296 (5577): 2395–8. doi:10.1126/science.1072068.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Unknown
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Unknown
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

File:T--SUSTech Shenzhen--tracking.mp4

The test of the CoChR using light with different wavelengths and intensity. The lights blue- and blue+ is from the projector whose wavelength is about 480 and the lights whose wavelengths are 395, 440, 470, 560 and 640 are from the LED of fluorescence microscope (Nikon eclipse Ti).

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 <partinfo>BBa_K2492001 SequenceAndFeatures</partinfo>
-[[File:T--SUSTech Shenzhen--cycle.gif|800px|thumb|left|GFP expressed in AWB| Control the C. elegans by the light: We used the optical fiber to form a light spot and the worm can follow the spot.]]
+[[File:T--SUSTech Shenzhen--tracking.mp4|800px|thumb|left| Control the C. elegans by the light: We used the optical fiber to form a light spot and the worm can follow the spot.]]
-[[File:T--SUSTech Shenzhen--getup.gif|800px|thumb|left|GFP expressed in AWB| The test of the CoChR using light with different wavelengths and intensity. The lights blue- and blue+ is from the projector whose wavelength is about 480 and the lights whose wavelengths are 395, 440, 470, 560 and 640 are from the LED of fluorescence microscope (Nikon eclipse Ti).]]
+[[File:T--SUSTech Shenzhen--getup.gif|800px|thumb|left| The test of the CoChR using light with different wavelengths and intensity. The lights blue- and blue+ is from the projector whose wavelength is about 480 and the lights whose wavelengths are 395, 440, 470, 560 and 640 are from the LED of fluorescence microscope (Nikon eclipse Ti).]]