Antiviral effect and application of epigenetic modification enzyme setd2
A technology for virus infection and hepatitis B virus, applied in the fields of biotechnology and medicine, can solve problems such as the unclear role of SETD2 protein, achieve the effect of preventing and inhibiting virus infection, wide application prospects, and improve survival rate
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Embodiment 1
[0107]Example 1: Interfering with the expression of SETD2 has a promoting effect on the replication of HBV in cells
[0108] HepG2 cells (purchased from ATCC) were cultured with DMEM medium, and the cells were transfected with HBV expression plasmids based on the framework of pHBV1.3, simulating the environment in which cells were infected with HBV. Then the cells were transfected with small interfering RNA against SETD2 (si-SETD2) and mock control (si-mimetic) (the transfection reagent INTERFERin was purchased from Polyplus).
[0109] The interfering RNA (Si-SETD2) against SETD2 and the mimic control (Si-mimetic) were purchased from Genephama Company, the sequence of Si-SETD2 is shown in SEQ ID NO: 5 and SEQ ID NO: 6, and the Si-mimetic The sequences are shown in SEQ ID NO: 7 and SEQ ID NO: 8. During synthesis, 2 dTs were added at the 3' to make the sequences more stable. The specific sequence is as follows:
[0110] Si-SETD2 sequence:
[0111] 5'-GGUGUAACUUAUGCAUUAATT-3' ...
Embodiment 2
[0120] Example 2: Overexpression of SETD2 inhibits HBsAg secretion in cells
[0121] Firstly, the cDNA of SETD2 (NM_014159.6) was divided into three segments and introduced into the eukaryotic expression vector pcDNA3.1 plasmid to construct the expression vectors of SETD2-F1, SETD2-F2 and SETD2-F3 respectively.
[0122] Wherein, the nucleotide sequence contained in SETD2-F1 is the 54th-2888th position of SEQ ID NO: 1, which encodes the 1st-945th position of human SETD2 protein SEQ ID NO: 2;
[0123] The nucleotide sequence contained in SETD2-F2 is SEQ ID NO: 1 2889-5267, which encodes human SETD2 protein SEQ ID NO: 2 946-1738, and this truncated body contains the SET domain;
[0124] The nucleotide sequence contained in SETD2-F3 is the 5268-7748 position of SEQ ID NO: 1, which encodes the 1739-2564 position of human SETD2 protein SEQ ID NO: 2, and this truncated body contains the WW domain.
[0125] SETD2-F1, SETD2-F2 and SETD2-F3 were respectively transfected into HepG2 cell...
Embodiment 3
[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 high-pressure tail vein, and 10ng pHBV1.3 plasmid was injected into the tail vein at the same time to simulate HBV infection in mice. After 72 hours, IFNα (15 mg / kg) was injected intraperitoneally. After 24 hours, the mouse liver tissue was dissected, and the expression of HBV core antigen (HBcAg) on the mouse liver was detected by immunofluorescence to indicate the HBV infection of the mouse.
[0132] The results of liver HBV-infected mice were as follows: Figure 4 shown.
[0133] The results showed that overexpression of SETD2-F2 in mice could inhibit HBV infection in mouse liver.
[0134] The results indicated that overexpression of SETD2 could inhibit virus infection in vivo.
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