Difference between revisions of "Part:BBa K3335005"

 
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<partinfo>BBa_K3335005 parameters</partinfo>
 
<partinfo>BBa_K3335005 parameters</partinfo>
 
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===CD47 siRNA overexpressed in HEK293T cell and exosome===
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<p>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.</p>
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https://2020.igem.org/wiki/images/1/17/T--NJU-China--7.png
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<p>Figure.1 CD47 siRNA is overexpressed is HEK293T cell</p>
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https://2020.igem.org/wiki/images/b/b2/T--NJU-China--8.png
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<p>Figure.2 CD47 siRNA is overexpressed in exosome</p>
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https://2020.igem.org/wiki/images/b/be/T--NJU-China--9.png
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<p>Figure.3 Absolute quantification of CD47 siRNA</p>
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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>
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<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>

Revision as of 11:58, 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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 42
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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.

T--NJU-China--7.png

Figure.1 CD47 siRNA is overexpressed is HEK293T cell

T--NJU-China--8.png

Figure.2 CD47 siRNA is overexpressed in exosome

T--NJU-China--9.png

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.