Difference between revisions of "Team:SUSTech Shenzhen/Project"
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<!-- 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. | <!-- 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. | ||
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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. | 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. | ||
Revision as of 03:31, 17 October 2016
Project Overview
test page
Contents
Abstract
Audiogenetics is a useful tool for high-efficiency cell regulation. Compared to chemical genetics, it stimulates cells with better precision and less toxicity. Furthermore, signals are conveyed to target cells with little delay, leading to a shorter response time.f(x) = \int_{-\infty}^\infty \hat f(\xi)\,e^{2 \pi i \xi x} \,d\xi
To achieve our goal of audiogenetics, membrane mechanosensitive channels (TRPC5 and Piezo) are chosen as receptors. Fluorescent calcium indicator (R-GECO) is employed to indicate cytoplasmic calcium level. Nuclear factor of activated T cells (NFAT) and YFP are used as downstream indicators to quantify the regulatory abilities.
f(x) = \int_{-\infty}^\infty \hat f(\xi)\,e^{2 \pi i \xi x} \,d\xi
Microfluidic channels are also utilized in the pre-study in which shear stress, similar to sound waves, is applied on cell surface as signal input to explore the basic parameters. A sound generator is constructed to test whether sound can trigger the channels as expected. Additionally, we use directed evolution to improve channels' selectivity for specific sound frequency and their sensitivity to sound of lower intensity.
| a | |
|---|---|
| DNA (500ng) | 8ul |
| Cutsmart buffer | 2.5ul |
| EcoRI-HF | 0.5ul |
| XbaI | 0.5ul |
| ddH2O | 16.5ul |
| Total | 25ul |
| msfGFP CDS① | msfGFP CDS② | |
|---|---|---|
| DNA(300ng) | 2.5ul | 2.5ul |
| Cutsmart buffer | 1ul | 1ul |
| MluI-HF/MscI | 0.5ul | 0.5ul |
| ddH2O | 6ul | 6ul |
| Total | 10ul | 10ul |
| a | |
|---|---|
| DNA (500ng) | 8ul |
| Cutsmart buffer | 2.5ul |
| EcoRI-HF | 0.5ul |
| XbaI | 0.5ul |
| ddH2O | 16.5ul |
| Total | 25ul |
Welcome to iGEM 2016!
Your team has been approved and you are ready to start the iGEM season!
Before you start
Please read the following pages:
Styling your wiki
You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.
While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.
Wiki template information
We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions">Pages for awards</a> link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!
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On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!
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Inspiration
You can also view other team wikis for inspiration! Here are some examples:
- <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a>
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- <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a>
- <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a>
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