Part:BBa_K2208004
A shRNA corresponding DNA sequence for BCL2 which could silence the gene.
This part is a short hairpin RNA (shRNA) sequence. When this sequence is cut by restriction enzyme and then integrated into pcDNA 6.2 vector, it can play an RNAi function in mammalian cell lines such as white fat adipose cells. When the shRNA vector loading anti-BCL2 is transfected into white fat adipose cells, the shRNA hairpin structure will be cleaved into anti-BCL2 siRNA by Dicer and then construct the RNA-induced silencing complex (RISC). The siRNA-RISC complex will target at BCL2 mRNA under the guide of siRNA sequence and digest it.
Usage and Biology
We packaged BCL2 siRNA into exosomes by transfecting HEK293 cells with a plasmid expressing BCL2 siRNA and then collected siRNA-encapsulated exosomes. When modified exosomes being intravenously injected, they will specifically recognize integrin receptors and fuse with white fat adipose cells under the direction of the tPep. Once inside cells, BCL2 siRNA will bind to BCL2 mRNA through base-pairing and digest the mRNA, resulting in sharp decrease of BCL2 protein in white fat adipose cells. As a consequence, BCL2 protein’s reduction and disturbed function will both result in the apoptosis of adipose cells, which ultimately have some therapeutic effects on losing weight.
CHARACTERIZATION
To ensure the interference efficiency of anti-BCL2 siRNA plasmid, we transfected it into HEK293 cells and then extracted protein from these cells to perform western blot. Significant down-regulation of BCL2 protein can be observed in HEK293 cells treated with anti-BCL2 siRNA, demonstrating that anti-BCL2 siRNA has the gene silencing effect. The siRNA-3 in the picture refers to this part RNA.
Figure 1. Protein quantitatively analysis of BCL2 protein after transfection of BCL2 siRNA
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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