Difference between revisions of "Part:BBa K3335005"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<p>At the same time, We detected siRNA in exosomes produced by HEK293T cells, confirmed that siRNA could be properly wrapped into exosomes, and prepared for its role (Figure.2).</p> | <p>At the same time, We detected siRNA in exosomes produced by HEK293T cells, confirmed that siRNA could be properly wrapped into exosomes, and prepared for its role (Figure.2).</p> | ||
<p>According to the absolute quantification of CD47 siRNA, we found that the amount of siRNA in HEK293T cells is 2.855E-08 nmol/L, the amount of siRNA in exosome is 2.544E-09 nmol/L. The exosome siRNA is equivalent to 8.9% of the siRNA concentration in the cell.</p> | <p>According to the absolute quantification of CD47 siRNA, we found that the amount of siRNA in HEK293T cells is 2.855E-08 nmol/L, the amount of siRNA in exosome is 2.544E-09 nmol/L. The exosome siRNA is equivalent to 8.9% of the siRNA concentration in the cell.</p> | ||
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| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K3335005 SequenceAndFeatures</partinfo> | ||
Revision as of 12:27, 22 October 2020
We used it to express iRGD-Lamp2b and siRNA targeted at CD47
Tumor cells escape macrophage phagocytosis by overexpressing CD47.Therefore, we combined the fusion expression of iRGD and Lamp2b to characterize the exosome targeting.At the same time, siRNA targeting CD47 was produced, and mRNA expression was targeted to be reduced.This allows macrophages to avoid the effects of CD47 and attack tumor cells.
CD47 siRNA overexpressed in HEK293T cell and exosome
First, we detected the correct expression of CD47 siRNA through RT-QPCR to ensure that sufficient targeted siRNA could be produced in cell. CMV-iRGD-siRC is this part. CMV-iRGD-siRP+C+K is a Composite part.
Figure.1 CD47 siRNA is overexpressed is HEK293T cell
Figure.2 CD47 siRNA is overexpressed in exosome
Figure.3 Absolute quantification of CD47 siRNA
At the same time, We detected siRNA in exosomes produced by HEK293T cells, confirmed that siRNA could be properly wrapped into exosomes, and prepared for its role (Figure.2).
According to the absolute quantification of CD47 siRNA, we found that the amount of siRNA in HEK293T cells is 2.855E-08 nmol/L, the amount of siRNA in exosome is 2.544E-09 nmol/L. The exosome siRNA is equivalent to 8.9% of the siRNA concentration in the cell.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 42
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]