The invention belongs to the technical field of
biomedicine, and particularly relates to
CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-Cas9) homing sequences and primers thereof, and a transgenic
expression vector and an establishment method thereof. The
CRISPR-Cas9 homing sequence sgRNA comprises SIDT1-
gene-specific
Target site-1 of which the target is positioned on E1
exon, and SIDT1-
gene-specific
Target site-2 of which the target is positioned on E2
exon, wherein the
nucleotide sequence of the
Target site-1 is disclosed as SEQ ID NO.8, and the
nucleotide sequence of the Target site-2 is disclosed as SEQ ID NO.9. The
CRISPR-Cas9 transgenic
expression vector is formed by connecting the Cas9 homing sequence Target site-1 and Target site-2 to the BsmB I and Bbs I
enzyme digestion sites of a recombinant shuttle Cas9 tool
plasmid. The recombinant shuttle Cas9 tool
plasmid can be quickly assembled with homing sequences of the two target sites of the
genome, thereby packaging the recombinant adenovirus vector specifically for the two target sites; and the recombinant shuttle Cas9 tool
plasmid can independently complete the editing of the large-segment
genome without dependence on the synergistic actions. The establishment process is simple and quick, and has high efficiency for performing the
genome large-segment editing function.