Difference between revisions of "Part:BBa K3335004"

Revision as of 11:53, 22 October 2020

We used it to express iRGD-Lamp2b and siRNA targeted at PD-L1

Tumor cells evade immune system surveillance by overexpressing PD-L1.Therefore, we combined the fusion expression of iRGD and Lamp2b to characterize the exosome targeting.At the same time, siRNA targeting PD-L1 is produced to reduce mRNA expression.This allows the immune system to avoid the effects of PD-L1 and attack tumor cells.

Sequence and Features

PD-L1 siRNA overexpressed in HEK293T cell and exosome

First, we detected the correct expression of PD-L1 siRNA through RT-QPCR to ensure that sufficient targeted siRNA could be produced in cell. CMV-iRGD-siR^P is this part. CMV-iRGD-siR^P+C+K is a Composite part.

Figure.1 PD-L1 siRNA is overexpressed is HEK293T cell

Figure2. PD-L1 siRNA is overexpressed in exosome

Figure.3 Absolute quantification of PD-L1 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 PD-L1 siRNA, we found that the amount of siRNA in HEK293T cells is 1.973E-08 nmol/L, the amount of siRNA in exosome is 4.370E-09 nmol/L. The exosome siRNA is equivalent to 22% of the siRNA concentration in the cell.