The invention relates to the technical field of
molecular biology and biological medicines, and particularly relates to application of gRNA sequences and combination thereof based on a
CRISPR system in treatment on
hepatitis B
virus. According to the method disclosed by the invention, eight types of guidance RNAs (gRNA) are designed according to design rules of
CRISPR gRNA and a conservative region of different genotypes of HBV sequences, and the eight types of guidance RNAs are structured on a PX330
expression vector. By utilizing the eight gRNAs in a
cell model, a mouse model and a
CRISPR / Cas9
system guided by combination of the
cell model and the mouse model, the expression and replication of the
hepatitis B
virus can be effectively inhibited. By united application of a plurality of gRNAs, a better effect can be achieved, and different genotypes of
HBV replication can be better inhibited. The
system has the characteristics of being easy to operate, high in inhibition efficiency on
HBV replication and applicable to various genotypes. Therefore, the gRNA and the combination thereof related to the invention are expected to be applied in preparation of a novel
drug for treating the
hepatitis B
virus.