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

Line 7: Line 7:
  
 
<body>
 
<body>
 +
    <br>
 +
    <br>
 +
    <br>
 
     <h1>Targeting Part</h1>
 
     <h1>Targeting Part</h1>
 
     <p>none</p>
 
     <p>none</p>
Line 15: Line 18:
 
     <h4>1)western blotting</h4>
 
     <h4>1)western blotting</h4>
 
     <p>Examine the function of KRAS siRNA with transfection into A549 cells using Lipo2000</p>
 
     <p>Examine the function of KRAS siRNA with transfection into A549 cells using Lipo2000</p>
−
     <h4>Evaluate the expression level of KRAS mRNA after transfected KRAS siRNA using Lipo 2000</h4>
+
     <h4>2)Evaluate the expression level of KRAS mRNA after transfected KRAS siRNA using Lipo 2000</h4>
 
     <br>
 
     <br>
 
     <h3>2.transfect KRAS siRNA and iRGD plasmid into HEK293 cells and collect exosomes</h3>
 
     <h3>2.transfect KRAS siRNA and iRGD plasmid into HEK293 cells and collect exosomes</h3>
 
     <br>
 
     <br>
 
     <h3>3.evaluate the effect of KRAS siRNA encapsulated by iRGD-modified exosomes on A549 cells</h3>
 
     <h3>3.evaluate the effect of KRAS siRNA encapsulated by iRGD-modified exosomes on A549 cells</h3>
−
     <h4>(1)western blotting</h4>
+
     <h4>1)western blotting</h4>
−
     <h4>qPCR</h4>
+
     <h4>2)qPCR</h4>
 +
    <br>
 
     <h3>4.cell proliferation assay</h3>
 
     <h3>4.cell proliferation assay</h3>
 
     <p> To ascertain that KRAS siRNA loaded into exosomes can suppress cancer cells Proliferation for further support</p>
 
     <p> To ascertain that KRAS siRNA loaded into exosomes can suppress cancer cells Proliferation for further support</p>
 
     <h3>5.transmission electron microscope assay</h3>
 
     <h3>5.transmission electron microscope assay</h3>
 
     <p> Observe the morphology of exosomes loaded with KRAS siRNA and determine density</p>
 
     <p> Observe the morphology of exosomes loaded with KRAS siRNA and determine density</p>
 +
    <br>
 
     <h3>6.Endotoxin detecting</h3>
 
     <h3>6.Endotoxin detecting</h3>
 
     <br>
 
     <br>

Revision as of 06:35, 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

2)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

2)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