Preliminary Study Of Lentiviral Vector-mediated CRISPR/Cas9 In The Treatment Of Rabies In Mice

Rabies virus(RABV)can cause viral encephalitis after infecting the host.It is an acute infectious disease virus,and the encephalitis is called rabies.Rabies is still one of the deadliest infectious diseases to humans,with a fatality rate close to 100%.Rabies exists in more than 60 countries,and most of the countries are developing countries.Current treatment methods still cannot successfully cure rabies.With the emergence and application of the CRISPR/Cas9 system,the lentiviral vector-mediated CRISPR/Cas9 system has been used to inhibit diseases caused by various viruses,and it also brings hope for the cure of rabies.Rabies virus N protein is highly conserved among different rabies virus strains and can interact with rabies virus polymerase to complete rabies virus replication and transcription.This study used CRISPR/Cas9 to knock out the N protein gene of the rabies virus,causing the N protein gene of the rabies virus to mutate and fail to function normally.It can effectively curb the replication of rabies virus at the cellular level and animal level(mice).The experiment predicted 2 pairs of sg RNA for the N protein,constructed two sets of Lenti CRISPR v2 vectors,and packaged and concentrated the lentivirus.First of all,experiments on mouse neurogenic cells found that concentrated lentivirus can significantly reduce the speed of rabies virus proliferation,and Lenti CRISPR-N277 lentivirus inhibits virus replication better than Lenti CRISPR-N394 lentivirus.Through a mouse model for challenge treatment experiments,it was found that the mice in the lentivirus injection group survived 5-7 days longer than the mice in the PBS control group.Whether on the level of cells infected with RABV or on the mouse model of RABV,the lentivirusmediated system can effectively inhibit rabies virus' s damage to the host,reduce virus replication at the cellular level,and extend mice at the animal level.Time to live.This study explored the use of lentiviral vectors to precisely knock out the N protein gene of rabies virus,inhibit the replication ability of rabies virus,and prolong the survival time of rabies-affected animals.