Difference between revisions of "Part:BBa K2492001"

Revision as of 15:22, 27 October 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.)

Sequence and Features

Assembly Compatibility:

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

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 <partinfo>BBa_K2492001 short</partinfo>
-Channelrhodopsins are a subfamily of retinylidene proteins (rhodopsins) that function as light-gated ion channels[1]. CoCHR is a kind of microbial-type channelrhodopsin originated from Chloromonas oogama, which is an unique ion channel served as light switch.In our case, CoCHR is a cation-selective light sensor, which promptly generates a large permeability of divalent cations after absorption of photon from our excitation source. In order to investigate how olfactory messages are transmitted by the neutron systems and activate AWA in certain purpose without much delay, we express CoCHR in AWA through linkage to the cell-specific promoter odr-10. Thus when we shed the ~400nm excitation light to transgenic C.elegans which exist the cell-specific promoter for AWA, the activated AWA is able to cause obvious response in body twisting comparing to wild type C.elegans. Whether the certain neuron is activated is reported by another part called GEM-GECO, a calcium indicated for excitation visualisation. In short, we establish a light-sensed AWA controlling system, so as to using this part to manipulate the activation of AWA, establish the response to input stimulation.
+The central part of our project. Part BBa_K2492001 contains CoCHR(Chloromonas oogama channelrhodopsin) [1].
-The merit of this part is sensitivity which is  five-times more sensitive to blue light than formal ChR2. Thus the low intensity requirement avoids the cross-activation of Crimson, which is another transmitted channelrhodopsin in AWB. Moreover, in order to confirm the where the target neurons are and roughly estimate the expression level of CoCHR, we also construct a fashion reporter protein called mcherry.
+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.)
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