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1. WO2022094730 - DNA CONSTRUCT FOR STABLY PRODUCING EMPTY CAPSIDS OF THE FOOT-AND-MOUTH DISEASE VIRUS IN MAMMALIAN CELLS; PROCESSES, USES, AND COMPOSITIONS THEREOF

Publication Number WO/2022/094730
Publication Date 12.05.2022
International Application No. PCT/CA2021/051592
International Filing Date 09.11.2021
IPC
C12N 15/86 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
86Viral vectors
A61K 39/135 2006.1
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
39Medicinal preparations containing antigens or antibodies
12Viral antigens
125Picornaviridae, e.g. calicivirus
135Foot-and-mouth disease virus
C07K 14/09 2006.1
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
005from viruses
08RNA viruses
085Picornaviridae, e.g. coxsackie virus, echovirus, enterovirus
09Foot-and-mouth disease virus
C12N 15/42 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
31Genes encoding microbial proteins, e.g. enterotoxins
33Genes encoding viral proteins
40Proteins from RNA viruses, e.g. flaviviruses
41Picornaviridae, e.g. rhinovirus, coxsackie viruses, echoviruses, enteroviruses
42Foot-and-mouth disease virus
C12N 15/56 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
52Genes encoding for enzymes or proenzymes
55Hydrolases (3)
56acting on glycosyl compounds (3.2), e.g. amylase, galactosidase, lysozyme
C12N 15/85 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
Applicants
  • INSTITUTO NACIONAL DE TECNOLOGIA AGROPECUARIA [AR]/[AR]
  • NATIONAL RESEARCH COUNCIL OF CANADA [CA]/[CA]
Inventors
  • MIGNAQUI, Ana Clara
  • WIGODOROVITZ, Andres
  • DUROCHER, Yves
Agents
  • BCF LLP
Priority Data
P2020010310209.11.2020AR
Publication Language English (en)
Filing Language English (EN)
Designated States
Title
(EN) DNA CONSTRUCT FOR STABLY PRODUCING EMPTY CAPSIDS OF THE FOOT-AND-MOUTH DISEASE VIRUS IN MAMMALIAN CELLS; PROCESSES, USES, AND COMPOSITIONS THEREOF
(FR) CONSTRUCTION D'ADN POUR PRODUIRE DE MANIÈRE STABLE DES CAPSIDES VIDES DU VIRUS DE LA FIÈVRE APHTEUSE DANS DES CELLULES DE MAMMIFÈRE ; PROCÉDÉS, UTILISATIONS ET COMPOSITIONS DE CELLE-CI
Abstract
(EN) The present invention refers to methods and compositions for increasing the production of large amounts of empty capsids of the foot-and-mouth disease virus (FMDV) in a stable manner in mammalian cells by regulating the expression of FMDV 3C protease. The instant methods and compositions are based on the fact that a decreased expression of 3C protease results in a reduced cell toxicity and an increased synthesis of viral capsid proteins, as well as production of recombinant empty capsids. The invention provides recombinant plasmids that direct the expression of P1, 3C, and the use of said plasmids for producing new stable cell lines capable of generating high titers of FMDV empty capsids. The invention provides methods for regulating the expression of the FMDV 3C protease gene at a transcriptional and translational level in order to achieve the required process level of 3C protease for the selection process, as well as the production process.
(FR) La présente invention concerne des procédés et des compositions permettant d'augmenter la production de grandes quantités de capsides vides du virus de la fièvre aphteuse (FMDV) de manière stable dans des cellules de mammifères en régulant l'expression de la protéase 3C du FMDV. Les procédés et compositions actuels sont basés sur la constatation qu'une diminution de l'expression de la protéase 3C entraîne une réduction de la toxicité cellulaire et une augmentation de la synthèse des protéines de capside virale, ainsi que la production de capsides vides recombinées. La présente invention concerne des plasmides recombinés commandant l'expression de P1, 3C, et l'utilisation desdits plasmides pour produire de nouvelles lignées cellulaires stables capables de générer des titres élevés de capsides vides de FMDV. La présente invention concerne également des procédés permettant de réguler l'expression du gène de la protéase 3C du virus de la fièvre aphteuse au niveau de la transcription et de la traduction afin d'atteindre le niveau de traitement requis de la protéase 3C pour le processus de sélection, ainsi que pour le processus de production.
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