Chinese Academy of Sciences

Fuqiang Xu

Yang Wu

Xiaobin He

Liangyu Jiang

Kunzhang Lin

Ting Mei

Zengpeng Han

A method of preparing a recombinant adeno-associated virus (rAAV) including: (1) preparing a shuttle plasmid and a corresponding recombinant bacmid including a baculovirus genome, where the shuttle plasmid includes at least an rAAV gene of interest flanked by inverted terminal repeats (ITR-GOI) integrated with a heterologous functional gene fragment, and the recombinant bacmid includes an expression cassette of functional protein components necessary for assembly of the rAAV; (2) integrating the rAAV ITR-GOI and the expression cassette of functional protein components by using the shuttle plasmid and the recombinant bacmid, to yield a recombinant bacmid including a recombinant baculovirus genome; and (3) transfecting, with the recombinant bacmid, a host cell line.

Method, system and recombinant bacmid for preparation of recombinant adeno-associated virus

A method for producing a recombinant adeno-associated virus (rAAV) and a recombinant baculovirus virus, the method including: (1) infecting an insect host packaging cell line with a corresponding recombinant baculovirus integrated with an rAAV genome ITR-GOI (gene of interest flanked by AAV inverted terminal repeats) and an AAV Cap gene or AAV Rep gene; (2) culturing the host packaging cell line infected with the recombinant baculovirus, so as to produce the recombinant adeno-associated virus; and (3) separating and purifying the recombinant adeno-associated virus obtained in (2).

Method of preparing recombinant adeno-associated virus and recombinant baculovirus

A method for producing recombinant adeno-associated virus, the method including: (1) transforming a gene of interest (GOI) into a recombinant baculovirus, where the recombinant baculovirus has a genome integrated with AAV Rep gene, Cap gene, and rAAV genome ITR-GOI that are needed in the production of the rAAV; and where the ITR-GOI is linked to expression cassette of the Cap gene and the Rep gene by a 5' terminal nucleic acid segment or a 3&#x2032; terminal nucleic acid segment; (2) infecting host insect larvae with the recombinant baculovirus prepared in (1), such that the rAAV is produced in vivo in the host insect larvae; and (3) lysing the host insect larvae obtained in (2), and extracting and purifying the rAAV.

Method for preparing recombinant adeno-associated virus

A recombinant baculovirus, including: an adeno-associated virus Rep gene, an adeno-associated virus Cap gene, and an recombinant adeno-associated virus genome ITR-GOI (gene of interest) flanked by rAAV inverted terminal repeats (ITR). The ITR-GOI includes a 5' terminal nucleic acid fragment and a 3&#x2032; terminal nucleic acid fragment. The ITR-GOI is linked to an expression cassette of the Cap gene and an expression cassette of the Rep gene through the 5&#x2032; terminal nucleic acid fragment and the 3&#x2032; terminal nucleic acid fragment, respectively.

Growing community of inventors

Wuhan, China

Fuqiang Xu