Title:
EXPRESSION CASSETTE FOR EXPRESSING GENE CONTAINING OVERLAPPING OPEN READING FRAME IN INSECT CELL, AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/039936
Kind Code:
A1
Abstract:
Provided are an expression cassette for expressing a gene containing an overlapping open reading frame in an insect cell, and the use thereof. The expression cassette contains, from 5' to 3', a promoter capable of driving transcription in an insect cell, an artificial construction sequence, and an overlapping open reading frame only lacking a first translation start codon, which are operably linked, wherein the artificial construction sequence contains a natural or artificially modified intron having a splicing activity in the insect cell, and the intron contains ATG, or the intron is located between any two adjacent nucleotides in ATG. Using a designed intron sequence, a recombinant adeno-associated virus vector containing the expression cassette regulates the relative expression of VP1, VP2 and VP3 proteins and the relative expression of Rep78 and Rep52 proteins by means of the splicing effect of the intron, and can mass produce rAAV with a high packaging rate and infectivity in an insect cell.
Inventors:
XIAO HE (CN)
HE XIAOBIN (CN)
HUANG GANG (CN)
HU YING (CN)
PAN XING (CN)
HUANG HE (CN)
DU LIANG (CN)
WANG MENGDIE (CN)
HE XIAOBIN (CN)
HUANG GANG (CN)
HU YING (CN)
PAN XING (CN)
HUANG HE (CN)
DU LIANG (CN)
WANG MENGDIE (CN)
Application Number:
PCT/CN2021/120828
Publication Date:
March 23, 2023
Filing Date:
September 27, 2021
Export Citation:
Assignee:
GENEVOYAGER WUHAN CO LTD (CN)
International Classes:
C12N15/866; C12N5/10; C12N7/01; C12R1/93
Domestic Patent References:
| WO2021113767A1 | 2021-06-10 |
Foreign References:
| CN113166731A | 2021-07-23 | |||
| CN102007209A | 2011-04-06 | |||
| CN101522903A | 2009-09-02 | |||
| CN111183225A | 2020-05-19 | |||
| CN106544325A | 2017-03-29 | |||
| CN101522903A | 2009-09-02 | |||
| CN103849629A | 2014-06-11 | |||
| CN112553257A | 2021-03-26 | |||
| CN112280801A | 2021-01-29 |
Other References:
CHEN, HAIFENG: "Intron Splicing-mediated Expression of AAV Rep and Cap Genes and Production of AAV Vectors in Insect Cells", MOLECULAR THERAPY, vol. 16, no. 5, 18 March 2008 (2008-03-18), pages 924 - 930, XP002586030, DOI: 10.1038/MT.2008.35
YLA-HERTTUALA S., MOL THER, vol. 20, 2012, pages 1831 - 1832
URABE ET AL., HUM GENE THER, vol. 13, 2002, pages 1935 - 1943
ASLANIDI ET AL., PROC. NATLACAD. SCI. USA, vol. 206, 2009, pages 5059 - 5064
YLA-HERTTUALA S., MOL THER, vol. 20, 2012, pages 1831 - 1832
URABE ET AL., HUM GENE THER, vol. 13, 2002, pages 1935 - 1943
ASLANIDI ET AL., PROC. NATLACAD. SCI. USA, vol. 206, 2009, pages 5059 - 5064
Attorney, Agent or Firm:
WUHAN HUAZHIYU INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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