The invention discloses a
cartilage cell targeting recombinant adeno-associated
virus and a preparation method thereof, and belongs to the technical field of
gene engineering. The recombinant adeno-associated
virus is obtained by co-transfecting an HEK293
cell by using a recombinant
plasmid pSSCMV-Cl2a1-SOX9 and a helper
plasmid; and the recombinant
plasmid pSSCMV-Cl2a1-SOX9 is obtained by inserting a
fusion gene of Col2a1 and SOX9 into a pSSHG-CMV carrier. The preparation method comprises the following steps: step 1, constructing a recombinant plasmid pSSCMV-Col2a1-SOX9, and constructing the recombinant plasmid pSSCMV-Col2a1-SOX9; step 2, culturing the HEK293 cells, and waiting for
transfection; and 3, co-transfecting the recombinant plasmid pSSCMV-Cl2a1-SOX9 obtained in the step 1, a pHelper plasmid and a pAAV-RC plasmid into the HEK293
cell cultured in the step 2, culturing, collecting a cell
lysis solution, and carrying out
iodixanol gradient centrifugation combined with
Heparin chromatographic purification to obtain the recombinant adeno-associated
virus. The recombinant adeno-associated virus obtained by the invention has targeting property and higher expression level.