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Publication Number
US-7524626-B2
Publication Date
2009-04-28
Expiration Date
Abstract
The invention includes methods of producing viral particles which include introducing into avian cells a nucleotide sequence encoding a replication deficient retroviral vector and introducing into the avian cells nucleotide sequences encoding products required for replication of the replication deficient retroviral vector under the control of a promoter that is functional in the avian cell line, and harvesting the viral particles.
Core Innovation
The invention relates to a method for transiently producing replication-deficient avian retroviral vectors by introducing into a cell a nucleotide sequence encoding an avian retroviral vector that is replication deficient, transiently introducing nucleotide sequences encoding the products required for replication of the replication deficient retroviral vector, the products being gag, pol and env proteins, and harvesting viral particles.
The approach is directed to producing replication-deficient avian retroviral vectors in an avian cell context, including an avian cell line, where the transiently introduced replication-related products comprise gag, pol and env proteins and the resulting viral particles are harvested for subsequent use.
The method further includes introducing the harvested viral particles into avian blastodermal cells and obtaining a transgenic avian species, thereby linking transient production of replication-deficient avian retroviral vector particles to generation of transgenic avian species.
Claims Coverage
The document includes three independent claims that collectively cover transient production of replication-deficient avian retroviral vector particles using gag, pol, and env replication products, and an application of the harvested particles for obtaining a transgenic avian species. Across these independent claims, the inventive coverage centers on transient introduction of the replication-deficient vector with replication-product nucleotide sequences followed by harvesting, with one claim further requiring introduction into avian blastodermal cells to obtain transgenic avians.
Transiently introducing replication-deficient avian retroviral vector and replication products with gag, pol and env proteins
A method comprising transiently introducing into a cell a nucleotide sequence encoding an avian retroviral vector wherein the avian retroviral vector is replication deficient; transiently introducing into the cell nucleotide sequence encoding products required for replication of the replication deficient retroviral vector, the products being gag, pol and env proteins; and harvesting viral particles.
Transiently introducing into an avian cell line replication-deficient avian retroviral vector and gag, pol and env replication products
A method comprising transiently introducing into an avian cell line a nucleotide sequence encoding an avian retroviral vector wherein the avian retroviral vector is replication deficient; transiently introducing into the avian cell line one or more nucleotide sequences wherein the nucleotide sequence(s) encode products required for replication of the replication deficient retroviral vector the products being gag, pol and env proteins; and harvesting viral particles.
Generating a transgenic avian species using harvested particles introduced into avian blastodermal cells
A method comprising introducing into a cell a nucleotide sequence encoding an avian retroviral vector wherein the avian retroviral vector is replication deficient; introducing into the cell nucleotide sequence wherein the nucleotide sequence encodes products required for replication of the replication deficient retroviral vector, the products being the gag, pol and env proteins; harvesting viral particles; introducing the harvested particles into avian blastodermal cells; and obtaining a transgenic avian species.
The independent claims cover transient production of replication-deficient avian retroviral vector particles by co-introducing an avian retroviral vector nucleotide sequence and gag, pol, and env replication-product nucleotide sequences into a cell or an avian cell line, followed by harvesting viral particles. One independent claim additionally requires introducing the harvested particles into avian blastodermal cells to obtain a transgenic avian species.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Obtaining a transgenic avian species by introducing harvested viral particles into avian blastodermal cells.
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