Study On Preventing Infecting Of RV Before Immune Response By RNAi

Rabies is a lethal human-and-animal infection disease caused by rabies virus. It spreaded by bitting and grabbing of infected animal, rabies virus approached nerve system of human-and-animal. Someone can suffered from by rabies though immune injecting on time. One of the factors of injecting is rabies virus reproducing and approaching nerve system before the body causing special neutralizing antibody. So preventing the reproducing of rabies virus before the immune response of vaccine is one of the key links to avoid being infected rabies after immune injecting.RNAi is post-transcriptional gene silencing induced by short-interfering RNA and founded in nematode firstly. That is to say a small duplex RNA binding with complementory mRNA in body can induce RNAi nuclease in cell degrading mRNA result in interrupting gene expressing.In this study, special siRNA can inhibit the reproducing of rabies virus by the inhibiting function of siRNA. Three transcription precursor of siRNA—small DNA molecular was designed according to the mRNA encoding glycoprotein and nucleoprotein of rabies virus. Then the six small DNA molecular was cloned into the vector of the psilencer2.1-V6, transcripted vector was acquired by indentifing sequence, then transfected into the cell of BHK-21 by stationary transfection and transient transfection. After aquiring stationary transfected cell line, detecting the reproduce of rabies virus using DFI and RT-PCR in 48h after inoculating rabies virus. The results show that the siRNA-N19 is prefer siRNA-G69 to inhibiting the reproducing of RV. The result is poor when detecting by DFI after stationary transfecting. We use the cell lysate of the BHK-G69 and BHK-N19 after stationary transfecting injecting the mouse inoculated RV, the fatal rate of the mouse is less than the mouse only injecting RV. It demonstrate that the special siRNA play a role on preventing the reproducing of RV in the body of the mouse.