Title:
EXPRESSION CASSETTE OF GENE CONTAINING OVERLAPPED OPEN READING FRAME AND APPLICATION OF EXPRESSION CASSETTE IN INSECT CELLS
Document Type and Number:
WIPO Patent Application WO/2023/092643
Kind Code:
A1
Abstract:
Provided are an expression cassette of a gene containing an overlapped open reading frame and an application of the expression cassette in insect cells. The expression cassette comprises: a promoter capable of driving transcription in the insect cells, an artificially constructed sequence, and an overlapped open reading frame only lacking a first translation initiation codon which are operatively connected from 5'- to 3'- terminal, wherein the artificially constructed sequence contains a first intron and a second intron which have splicing activity in the insect cells, the first intron and the second intron share a 5' part or a 3' part, and the artificially constructed sequence does not contain a translation initiation codon; and in a post-transcription processing process, a translation initiation codon AUG can be formed by means of the alternative splicing effect of the first intron and the second intron. The relative expression of different capsid proteins and different Rep proteins is regulated and controlled by using the alternative splicing effect of the introns, and rAAV with high packaging rate and infectivity can be produced in the insect cells on a large scale.
Inventors:
XIAO HE (CN)
HE XIAOBIN (CN)
HUANG GANG (CN)
HU YING (CN)
DU LIANG (CN)
WANG MENGDIE (CN)
HE XIAOBIN (CN)
HUANG GANG (CN)
HU YING (CN)
DU LIANG (CN)
WANG MENGDIE (CN)
Application Number:
PCT/CN2021/135642
Publication Date:
June 01, 2023
Filing Date:
December 06, 2021
Export Citation:
Assignee:
GENEVOYAGER WUHAN CO LTD (CN)
International Classes:
C12N15/866; C07K14/015; C12N5/10; C12N15/35
Domestic Patent References:
| WO2021077017A1 | 2021-04-22 |
Foreign References:
| CN101522903A | 2009-09-02 | |||
| US20100233685A1 | 2010-09-16 | |||
| CN101213203A | 2008-07-02 | |||
| CN111183225A | 2020-05-19 | |||
| CN101421408A | 2009-04-29 | |||
| CN106544325A | 2017-03-29 | |||
| CN101522903A | 2009-09-02 | |||
| CN103849629A | 2014-06-11 | |||
| CN112553257A | 2021-03-26 | |||
| CN112280801A | 2021-01-29 |
Other References:
NIELSEN HENRIK; WERNERSSON RASMUS: "An overabundance of phase 0 introns immediately after the start codon in eukaryotic genes", BMC GENOMICS, vol. 7, no. 1, 11 October 2006 (2006-10-11), London, UK , pages 1 - 15, XP021022221, ISSN: 1471-2164, DOI: 10.1186/1471-2164-7-256
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, 1 May 2008 (2008-05-01), GB , pages 924 - 930, XP002586030, ISSN: 1525-0024, 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
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, 1 May 2008 (2008-05-01), GB , pages 924 - 930, XP002586030, ISSN: 1525-0024, 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
Attorney, Agent or Firm:
WUHAN HUAZHIYU INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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