Difference between revisions of "Team:LZU-CHINA/Futurework"

1.Test the effectiveness of 5HRE-miniCMV promoter.

We will first construct 5HRE-miniCMV promoter, and then construct 5HRE-miniCMV-EGFP lentiviral plasmid. Use CoCl2-6H2O at different concentrations to induce hypoxia condition of 293T cell lines, and EGFP expression will be observed under an inverted fluorescence microscope.

2.Test the anti-tumor activity of other miRNA besides mir-135b-3p.

We will construct stable cell lines to overexpress mir-942-5p and mir-769-5p in MKN-45 and SGC7901 cell lines. Flow cytometry, scratch assay and CCK8 assay was respectively used to verify the inhibitory effect of microRNAs on gastric cancer cells at the cellular biological level.

3.Test the effectiveness of BS-Biotin-On system.

We will constrcut BS-Biotin-On-EGFP plamid to verify the effectiveness of BS-Biotin-On system and then construct regulatory and response vectors to control the expression of downstream interest gene (mir-135b-3p).

4.Test the anti-tumor activity of other miRNA besides mir-135b-3p.

We will construct stable cell lines to overexpress mir-942-5p and mir-769-5p in MKN-45 and SGC7901 cell lines. Flow cytometry, scratch assay and CCK8 assay was respectively used to verify the inhibitory effect of microRNAs on gastric cancer cells at the cellular biological level.

5.Identify target genes of miRNA.

Using RNA-seq and dual luciferase reporter system to identify target genes of miRNA.

6.Test the effectiveness of ECOB-P

minCMV

and P

GAL1

We will constrcut ECOB-P

minCMV

and P

GAL1

plamids to verify whether tetracycline and galactosethe can respectively induce the expression of downstream interest gene mir-942-5p and mir-769-5p.

7. Fuse all our modules in one plasmid

To explore the best concentration of inducers that have the most effective anti-tumor activity.

8.Replace 293T cells with TIL cells.

Excise TIL cells from patients’(gastric cancer) body and then repeat all our experiments at the level of TIL cells.