Antiviral effect and application of epigenetic modifying enzyme SETD2
A virus infection and virus infection-related technology, applied in the field of epigenetic modification enzyme SETD2, can solve the problem of unclear role of SETD2 protein, and achieve the effect of preventing and inhibiting virus infection, inhibiting virus infection and improving curative effect.
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[0107] Example 1: Interference with the expression of SETD2 can promote HBV replication in cells
[0108] HepG2 cells (purchased from ATCC) were cultured in DMEM medium, and the cells were transfected with an HBV expression plasmid based on the framework of pHBV1.3 to simulate the environment of cell infection with HBV. Then the cells were transfected with small interfering RNA for SETD2 (si-SETD2) and a mock control (si-mock) (transfection reagent INTERFERin was purchased from Polyplus).
[0109] Interfering RNA for SETD2 (Si-SETD2) and mimetic control (Si-mimetic) were purchased from Genephama. The sequence of Si-SETD2 is shown in SEQ ID NO: 5 and SEQ ID NO: 6. The sequence is as shown in SEQ ID NO: 7 and SEQ ID NO: 8, adding 2 dT at 3'to make the sequence more stable. The specific sequence is as follows:
[0110] Si-SETD2 sequence:
[0111] 5'-GGUGUAACUUAUGCAUUAATT-3' (Shunyi);
[0112] 5'-UUAAUGCAUAAGUUACACCTT-3' (antisense).
[0113] Si-simulant sequence:
[0114] 5'-UUCUCCGAACGUG...
Example Embodiment
[0120] Example 2: Overexpression of SETD2 inhibits HBsAg secretion in cells
[0121] First, the cDNA of SETD2 (NM_014159.6) was divided into three segments and introduced into the eukaryotic expression vector pcDNA3.1 plasmid, respectively, to construct SETD2-F1, SETD2-F2, and SETD2-F3 expression vectors.
[0122] Wherein, the nucleotide sequence contained in SETD2-F1 is SEQ ID NO: 1 54-2888, encoding human SETD2 protein SEQ ID NO: 2 1-945;
[0123] SETD2-F2 contains the nucleotide sequence of SEQ ID NO:1, positions 2889-5267, encoding human SETD2 protein SEQ ID NO: 2, positions 946-1738, and this truncated body contains the SET domain;
[0124] The nucleotide sequence contained in SETD2-F3 is SEQ ID NO:1 5268-7748, encoding human SETD2 protein SEQ ID NO: 2 1739-2564, and this truncated body contains a WW domain.
[0125] SETD2-F1, SETD2-F2, and SETD2-F3 were transfected into HepG2 cells at a density of 1ng / ml, and pHBV1.3 plasmid was transfected simultaneously. After 48 hours, replace...
Example Embodiment
[0130] Example 3: Overexpression of SETD2 inhibits HBV infection in mouse liver
[0131] Construct SETD2-F2 expression vector (same as Example 2). 20ng SETD2-F2 was injected into mice (8-week-old male SDF grade C57BL6 mice, purchased from Sipper BK) through tail vein high pressure injection, and 10ng pHBV1.3 plasmid was injected with tail vein high pressure at the same time to simulate HBV infection in mice. IFNα (15 mg / kg) was injected intraperitoneally 72 hours later. Dissected after 24 hours, took mouse liver tissue, and used immunofluorescence method to detect the expression of HBV core antigen (HBcAg) on the mouse liver to indicate the HBV infection of the mouse.
[0132] The results of liver HBV infection in mice Figure 4 Shown.
[0133] The results showed that overexpression of SETD2-F2 in mice can inhibit HBV infection in the liver of mice.
[0134] The results indicate that overexpression of SETD2 can inhibit virus infection in vivo.
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