Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1990-06-25
1993-10-12
Schwartz, Richard A.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
4351722, 4351723, 435239, 4352401, 4352402, 4353201, 935 24, 935 32, 935 34, 935 57, 935 70, C12N 1586, C12N 510, C12N 701, C12P 2100
Patent
active
052524655
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to new viral vectors for the integration and expression of a heterologous gene in avian cells, and constitutes a development of the invention which is the subject of Patent Applications FR 84/15,764 and EP 178,996 (85 40 1999), to which reference should be made for a better understanding of the present application.
The present invention also relates to plasmids for the integration and expression of a heterologous gene in competent cells.
The present invention relates, in addition, to cells infected or transfected by these viral or plasmid vectors.
Finally, the present invention relates to a method for the production of a protein by culturing these infected or transfected cells, as well as to a method for the genetic modification of animals using these viral vectors.
The retroviral genome represents an integrated structure which drives a cyclic process; one of the phases of this process comprises a regular integration of the viral genome in the cell DNA. This feature is associated with other properties which, by various mechanisms, endow some of these viruses with pathogenic properties with respect to their hosts.
An objective of the present invention is to use certain properties of retroviral structures for transferring genes to whole animals, in their somatic structures for the purpose of "gene therapy", or in their germinal structures for the purpose of transmission to the progeny. To achieve this objective, it is necessary to break up the process of operation of the retroviral structure in order to retain only the component of it whose use is justified by the desired objective. At the same time, all the parts liable to exert pathogenic effects will be eliminated. In particular, all transmission to the target subjects of potentially dangerous oncogenes, or of structures capable of permitting viral multiplication to continue in the chosen target, will have to be absolutely prohibited.
An object of the invention is, in effect, to obtain viral vectors capable of a high transfer efficiency which are usable for infecting the animal and, to this end, to obtain the situation where the viruses, actively multiplied in vitro, are capable of being inserted into the animal but become incapable of multiplying therein.
An object of the present invention is hence to produce vectors relieved of oncogenic sequences and capable of transporting other genes, in a configuration permitting the most efficacious possible expression of the latter.
Thus, in EP 178,996, the main vectors described still contained the erbA oncogene. According to the present invention, this oncogene has, in its turn, been deleted.
According to the present invention, optimization of retroviral vectors, the subject of the previous patents cited above, was sought.
Optimization of the vectors was sought by working, respectively, on the activity of the promoters and on the efficacy of the system of maturation and translation of the RNAs.
The subject of the present invention is, in effect, a viral vector for the integration and expression of at least one heterologous gene in avian cells, which consists wholly or in part of the proviral genome of avian erythroblastosis or of a related virus in which said heterologous gene(s) replace(s) the v-erbA gene and/or the v-erbB gene, and wherein said gene(s) is/are either controlled by an LTR promoter of the same virus, in which case the heterologous gene(s) mimic(s) the gene(s) it/they replace(s), or controlled by a heterologous promoter, in which case an additional att sequence is situated upstream from said heterologous promoter.
Heterologous gene is understood to denote a gene which is not normally present in the genome of the avian erythroblastosis virus. It can be a so-called "useful" gene coding for a protein of industrial interest intended for production by cell culturing, or alternatively a gene which can be of special interest in the treatment or breeding of birds, in particular chickens, a gene providing for vaccination or better development, for example, or it can alternativ
REFERENCES:
patent: 4957865 (1990-09-01), Samarut et al.
Bagnis Claude
Belakebi Mustapha
Benchaibi Miloud
Chebloune Yahia
Cosset Francois-Loic
Institut National de la Recherche Agronomique (IMPA)
Railey II Johnny F.
Schwartz Richard A.
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