Description: We are not satisfied to use the traditional ways to regulate gene expression, but to choose auodiogenetics and optogenetics to establish a high-efficiency regulation platform. Compared to the chemogenetics, sound and light are precise and nontoxic to our objects. Furthermore, the signals are easy to input into the system without delay.
 We try to use a membrane mechanosensitive channel- transient receptor potential channel (TRPC5) to transform the audio wave energy as the input into cho-k1 cell. To select the cells which are sensitive to the specific wavelength, we mutate TRPC5 by molecular engineering and use calcium indicator(R-GECO) to quantify the intracellular calcium. Finally, we use the YFP as the downstream output signal to quantitatively analysis the regulatory ability of audiogenetics. At the same time, we attempt to use opsin chrimson and coch-R expressed in C.elegans neurons, exploring a new way to study the neuronal response and learning process of worms. We choose a pair of sensor neurons AWA\AWB which are respectably and directly related to attractive and repulsive odorants. By using light-ray instead of chemicals, we can manufacture the alternative and consistent neuronal stimulus and observe its neuronal pathway in vivo. Microfluidics is an important tool used in our experiments. Cells or C.elegans are seeded in the channels of microfluidics undergoing shear force controlled by microfluidics apparatus. We utilize shear stress as mechanical wave to impose pressure on cell membrane by changing the velocity of liquid flow in channels of microfluidics. 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.
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Year: 2016Visit Wiki
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Updated at: 8/9/16