Difference between revisions of "Team:SUSTech Shenzhen"

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Another technology used is directed evolution. Here, we apply random PCR to acquire a large library of mutants of mechanosensitive channels which are then screened and selected based on the downstream pathways we design to meet our needs.  -->
 
Another technology used is directed evolution. Here, we apply random PCR to acquire a large library of mutants of mechanosensitive channels which are then screened and selected based on the downstream pathways we design to meet our needs.  -->
  
<p>Auodiogenetics is a good tool to establish a high-efficiency regulation platform. Compared to the chemogenetics, sound is more precise and nontoxic to cells. Furthermore, it is easy to convey the signal into cells without delay. To realize it, we use membrane mechxnosensitive channels(TRPC5 and Piezo)as receptors. Then we employ calcium indicator to reveal the level of intracellular calcium ions. Finally, we use the nuclear factor of activated T cells(NFAT) and YFP as downstream to quantify the regulatory ability. Microfluidics is also used in our experiments. We utilize shear stress as a mechanical force which is similar to sound for the primary examination. Then, we use sound generator to demonstrate if sound is able to trigger the channel also. Considering that normal channel may not be sensitive enough to sound of normal intensity, also to improve channel's specificity for special wavelength, directed evolution is used to enhance channel’s sensibility and specificity.</p>
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<p>Audiogenetics is a good tool to establish a high-efficiency regulation platform. Compared to chemogenetics, sound is more precise and nontoxic to cells. Furthermore, it is easy to convey the signal into cells without delay. To realize it, we use membrane mechanosensitive channels(TRPC5 and Piezo)as receptors. Then we employ fluorescent calcium indicator(R-GECO) to reveal the level of intracellular calcium ions. Finally, we use nuclear factor of activated T cells(NFAT) and YFP as downstream indicators to quantify the regulatory abilities of mechanosensitive channels. Microfluidics is also used in our experiments. We utilize shear stress as a mechanical force which is similar to sound in the initial examination. Then, we use sound generator to demonstrate if sound is able to trigger the channels as well. To improve channel's specificity for special wavelength and sensitivity to sound of normal intensity, directed evolution is used in our project.</p>
 
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Revision as of 16:21, 15 July 2016

Abstract

Audiogenetics is a good tool to establish a high-efficiency regulation platform. Compared to chemogenetics, sound is more precise and nontoxic to cells. Furthermore, it is easy to convey the signal into cells without delay. To realize it, we use membrane mechanosensitive channels(TRPC5 and Piezo)as receptors. Then we employ fluorescent calcium indicator(R-GECO) to reveal the level of intracellular calcium ions. Finally, we use nuclear factor of activated T cells(NFAT) and YFP as downstream indicators to quantify the regulatory abilities of mechanosensitive channels. Microfluidics is also used in our experiments. We utilize shear stress as a mechanical force which is similar to sound in the initial examination. Then, we use sound generator to demonstrate if sound is able to trigger the channels as well. To improve channel's specificity for special wavelength and sensitivity to sound of normal intensity, directed evolution is used in our project.

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