Application of miR-199a-3p in preparation of medicine for resisting hepatitis B virus
A mir-199a-3p, 1.mir-199a-3p technology, applied in antiviral agents, drug combinations, pharmaceutical formulations, etc., can solve problems such as incomplete elucidation of the mechanism of action
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[0023] 1. Cell Culture
[0024] HepG2 2.2.15 and HEK293 cell lines were cultured in DMEM and MEM-α (GIBCO, USA) basal medium respectively, both mediums were supplemented with 10% fetal bovine serum, 20mM HEPES, 2mM glutamine, 100 units / ml penicillin and 100 μg / ml streptomycin, all cells were placed in 5% CO 2 In a 37°C incubator, subculture every 2-3 days.
[0025] 2. Cell Transfection
[0026] The 2'methoxy (2'-OMe) modified antisense oligonucleotide (ASO) that is completely complementary to human mature miRNA was synthesized by IDT, and the cells were prepared in 10 4Each well was inoculated in a 96-well plate, and the next day, the cells were transfected with 50nM miRNA 2'-OMe-ASO or 5ng / μL miRNA expression vector with Lipofectamine 2000, and each group of experiments was done in 3 replicate wells, according to the laboratory Conventional methods and refer to instructions. After 48 hours, the culture medium was changed, and after 24 hours, the cell supernatant was coll...
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