Team:NJU-China/Attributions
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
