US 11,708,585 B2 · Method, system and recombinant bacmid for preparation of recombinant adeno-associated virus

Abstract

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.

Claims6

Dependent claims

A preparation system of rAAV, comprising a shuttle plasmid and a first recombinant bacmid;

wherein:
(i) the shuttle plasmid comprises an ITR-GOI, and an expression cassette of a Cap gene of AAV;
and
the first recombinant bacmid comprises a baculovirus genome having non-essential genes of Chia, and Cath absent, and an expression cassette of a Rep gene of AAV;
wherein the expression cassette of the Rep gene of AAV is inserted into loci of the absent Chia and Cath genes;
or
(ii) the shuttle plasmid comprises the ITR-GOI, and an expression cassette of a Rep gene of AAV;
and
the first recombinant bacmid comprises a baculovirus genome having non-essential genes of Chia, and Cath absent, and an expression cassette of a Cap gene of AAV;
wherein the expression cassette of the Cap gene of AAV is inserted into loci of the absent Chia and Cath genes.

7 more elements

Dependencies are inferred from the claim text.

Facts

Application

16/455,745Filed 28 Jun 2019 · continuation in part

Priority

CN 201811618542.X28 Dec 2018

Published

US 2020/0208175 A12 Jul 2020

Granted

25 Jul 2023

Term ends

Not estimatedThe source records don’t support an estimate.

Classification13 codes

C12N 15/86
For animal cells: viral vectors
C07K 14/005
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof: from viruses
C12N 2710/10043
Use of virus, viral particle or viral elements as a vector: viral genome or elements thereof as genetic vector
C12N 2710/10052
Methods of production or purification of viral material: relating to complementing cells and packaging systems for producing virus or viral particles

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

Life of this patent

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

Life of this patent

Abstract

Claims6