Difference between revisions of "Part:BBa K2989002"

Revision as of 12:22, 21 October 2019

miR-HBsAg-70

MiRNAs are a class of evolutionarily highly conserved single-stranded, non-coding small RNAs that are about 18 to 25 nt long. This kind of RNAs play important roles in many endogenic processes. Mostly, it functions as a post-transcriptional regulator of gene expression, which usually results in the silence of target gene. This process is commonly known as gene silencing.

Usage

Our miR-HBsAg-70 can function at the 70bp of "Trojan horse" fragment (92 amino acid length HBsAg fragment).,and therefore, leads to the silence of our "Trojan horse" fragment .In this case, we can ensure that "Trojan horse" fragment will not be expressed in target cells, since the gene will be silenced by our part.

A: pCDNA6.2-hTERT- HBsAg EmGFP-miR-Control
B: pCDNA6.2-hTERT-HBsAg-EmGFP-miR-70
We constructed pCDNA6.2-hTERT-HBsAg-EmGFP-miR-70 and transfected the element plasmid into HepG2 cells, and observed EmGFP expression by fluorescence microscopy after 36 h. All EmGFPs were taken with the same parameter settings to ensure the comparability of the experimental results. It can be seen from the figure that compared to Figure A (pCDNA6.2-hTERT-HBsAg-EmGFP-miR-Control plasmid) without miRNA elements, Figure B showed a relatively faint green fluorescence intensity, demonstrating that the miRNA exerts an inhibitory regulatory effect on the target sequence. We verified that the part can work. Sequence and Features