Difference between revisions of "Team:NJU-China/Attributions"

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     <h3>Targeting Part</h3>
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    <p>none</p>
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    <br>
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                <div class="nav-wrapper"><a class="page-title">Attributions</a></div>
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    <h3>Silencing Part</h3>
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            </div>
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    <h4>in vitro</h4>
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    <h5>1.examine the function of one basic part: KRAS siRNA</h5>
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        <div class="container">
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    <h6>1)western blotting</h6>
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            <a href="#" data-activates="nav-mobile" class="button-collapse top-nav full hide-on-large-only"> <i class="material-icons">menu</i> </a>
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    <p>Examine the function of KRAS siRNA with transfection into A549 cells using Lipo2000</p>
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    <h6>Evaluate the expression level of KRAS mRNA after transfected KRAS siRNA using Lipo 2000</h6>
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    <br>
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    <h5>2.transfect KRAS siRNA and iRGD plasmid into HEK293 cells and collect exosomes</h5>
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                <img class="background responsive-img" src="https://static.igem.org/mediawiki/2016/c/c8/NJU_China_2016_iGEM_logo.png" alt="NJU-China LOGO"> </li>
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    <br>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Background" class="waves-effect waves-teal">Background</a></li>
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    <h5>3.evaluate the effect of KRAS siRNA encapsulated by iRGD-modified exosomes on A549 cells</h5>
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    <h6>(1)western blotting</h6>
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    <h6>qPCR</h6>
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                    <li class="bold"> <a class="collapsible-header waves-effect waves-teal">Project</a>
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    <h5>4.cell proliferation assay</h5>
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    <p> To ascertain that KRAS siRNA loaded into exosomes can suppress cancer cells Proliferation for further support</p>
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                            <ul>
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    <h5>5.transmission electron microscope assay</h5>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project#Design">Design</a></li>
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    <p> Observe the morphology of exosomes loaded with KRAS siRNA and determine density</p>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project#Results">Results</a></li>
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    <h5>6.Endotoxin detecting</h5>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project#Conclusion">Conclusion</a></li>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Project#Future_Work">Future Work</a></li>
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                            </ul>
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    <h4>in vivo</h4>
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    <h5>1.establish the mouse model</h5>
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    <p>Injecte mice subcutaneously with A549-LUC cells</p>
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    <br>
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    <h5>2.treatment</h5>
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    <p>randomly assign the mice into 2 groups and treat with different injections: PBS and iRGD-modified exosomes loaded with KRAS siRNA</p>
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                <ul class="collapsible collapsible-accordion">
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                    <li class="bold"> <a class="collapsible-header waves-effect waves-teal">Model</a>
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     <h5>3.collect data</h5>
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    <p>Kill the mice and harvest tumors, compare the weight and volumes.</p>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Model#Delivery_Model">Delivery Model</a></li>
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     <h5>4.western blotting</h5>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Model#RNAi_Model">RNAi Model</a></li>
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     <p> extract protein from tumor issues and evaluate the K-ras protein expression Changes</p>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Model#Signaling_Model">Signaling Model</a></li>
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     <h5>5.qPCR</h5>
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     <p>extract RNA from tumor issues and evaluate the expression level of KRAS mRNA</p>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Parts" class="waves-effect waves-teal">Parts</a></li>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Safety" class="waves-effect waves-teal">Safety</a></li>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Human_Practice" class="waves-effect waves-teal">Human Practice</a></li>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Team" class="waves-effect waves-teal">Team</a></li>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Attributions" class="waves-effect waves-teal">Attributions</a></li>
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            <li class="bold"><a href="https://2016.igem.org/Team:NJU-China/Collaborations" class="waves-effect waves-teal">Collaborations</a></li>
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                    <li class="bold"> <a class="collapsible-header waves-effect waves-teal">Notebook</a>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Notebook#Methods">Methods</a></li>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Notebook#Protocol">Protocol</a></li>
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                                <li><a href="https://2016.igem.org/Team:NJU-China/Notebook#Notebook">Notebook</a></li>
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                <p>Abcdefg hijklmnop qrs tuv wx yz. Abcdefg hijklmnop qrs tuv wx yz. Abcdefg hijklmnop qrs tuv wx yz. Abcdefg hijklmnop qrs tuv wx yz. Abcdefg hijklmnop qrs tuv wx yz. Abcdefg hijklmnop qrs tuv wx yz. Abcdefg hijklmnop qrs tuv wx yz. Abcdefg</p>
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Revision as of 06:33, 13 October 2016

Targeting Part

none


Silencing Part

in vitro

1.examine the function of one basic part: KRAS siRNA
1)western blotting

Examine the function of KRAS siRNA with transfection into A549 cells using Lipo2000

Evaluate the expression level of KRAS mRNA after transfected KRAS siRNA using Lipo 2000

2.transfect KRAS siRNA and iRGD plasmid into HEK293 cells and collect exosomes

3.evaluate the effect of KRAS siRNA encapsulated by iRGD-modified exosomes on A549 cells
(1)western blotting
qPCR
4.cell proliferation assay

To ascertain that KRAS siRNA loaded into exosomes can suppress cancer cells Proliferation for further support

5.transmission electron microscope assay

Observe the morphology of exosomes loaded with KRAS siRNA and determine density

6.Endotoxin detecting


in vivo

1.establish the mouse model

Injecte mice subcutaneously with A549-LUC cells


2.treatment

randomly assign the mice into 2 groups and treat with different injections: PBS and iRGD-modified exosomes loaded with KRAS siRNA


3.collect data

Kill the mice and harvest tumors, compare the weight and volumes.


4.western blotting

extract protein from tumor issues and evaluate the K-ras protein expression Changes


5.qPCR

extract RNA from tumor issues and evaluate the expression level of KRAS mRNA