Complement-resistant non-mammalian DNA viruses and uses thereof
AssigneesThe General Hospital Corporation, Biogen, Inc.
InventorsFrederick M. Boyce, James G. Barsoum
Life of this patent
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Abstract
Disclosed are methods, nucleic acids, and cells for expressing an exogenous gene in a mammalian cell, involving (i) introducing into the cell a complement-resistant non-mammalian DNA virus (e.g., a baculovirus), optionally having an altered coat protein, the genome of which virus carries an exogenous gene, and (ii) growing the cell under conditions such that the gene is expressed.
Claims45
A method for producing a complement-resistant non-mammalian DNA virus, the method comprising:
- introducing into an Estigmene acrea cell a genome of a non-mammalian DNA virus selected from the group consisting of baculoviruses, entomopox viruses, and densonucleosis viruses, wherein the genome comprises an exogenous gene operably linked to a mammalian-active promoter;
- and
- allowing the virus to replicate in the Estigmene acrea cell, thereby producing a complement-resistant non-mammalian DNA virus.
A method for producing a complement-resistant non-mammalian DNA virus, the method comprising:
- providing a non-mammalian cell that expresses one or both of (i) a mammalian siayltransferase and (ii) a mammalian galactosyltransferase;
- introducing into the cell a non-mammalian DNA virus, wherein the genome of the virus comprises an exogenous gene operably linked to a mammalian-active promoter;
- and
- allowing the virus to replicate in the non-mammalian cell, thereby producing a complement-resistant non-mammalian DNA virus.
A method for producing a complement-resistant non-mammalian DNA virus, the method comprising:
- introducing into a non-mammalian cell a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises an exogenous gene operably linked to a mammalian-active promoter;
- culturing the non-mammalian cell in a culture medium comprising one or both of (i) D-mannosamine and (ii) N-acetyl-D-mannosamine;
- and
- allowing the virus to replicate in the non-mammalian cell, thereby producing a complement-resistant non-mammalian DNA virus.
A method for producing a complement-resistant non-mammalian DNA virus, the method comprising:
- introducing into a non-mammalian cell a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises
- (i) an exogenous gene operably linked to a mammalian-active promoter and
- (ii) one or both or (a) a mammalian siayltransferase gene and (b) a mammalian galactosyltransferase gene, wherein the siayltransferase and/or galactosyltransferase gene is operably linked to a promoter that is active in the non-mammalian cell;
- and
- allowing the virus to replicate in the non-mammalian cell, thereby producing a complement-resistant non-mammalian DNA virus.
A method for producing a complement-resistant non-mammalian DNA virus, the method comprising:
- providing a non-mammalian cell that expresses one or both of (i) a CD59, or a homolog thereof and (ii) a decay accelerating factor (DAF), or a homolog thereof;
- introducing into the cell a non-mammalian DNA virus, wherein the genome of the virus comprises an exogenous gene under the control of a mammalian-active promoter;
- and
- allowing the virus to replicate in the non-mammalian cell, thereby producing a complement-resistant non-mammalian DNA virus.
A method for producing a complement-resistant non-mammalian DNA virus, the method comprising:
- introducing into a non-mammalian cell a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises
- (i) an exogenous gene operably linked to a mammalian-active promoter and
- (ii) one or both or
- (a) a nucleotide sequence encoding CD59, or a homolog thereof, operably linked to a promoter that is active in the non-mammalian cell and
- (b) a nucleotide sequence encoding decay accelerating factor, or a homolog thereof, operably linked to is a promoter that is active in the non-mammalian cell;
- and
- allowing the virus to replicate in the non-mammalian cell, thereby producing a complement-resistant non-mammalian DNA virus.
An Estigmena acrea cell comprising a genome of a non-mammalian DNA virus selected from the group consisting of baculoviruses, entomopox viruses, and densonucleosis viruses, wherein the genome comprises an exogenous gene under the control of a mammalian-active promoter.
A non-mammalian cell comprising
- (i) a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises an exogenous gene under the control of a mammalian-active promoter and
- (ii) one or both of (a) a nucleic acid sequence encoding a mammalian siayltransferase and (b) a nucleic acid sequence encoding a mammalian galactosyltransferase.
A cell culture comprising:
- (i) a non-mammalian cell containing a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises an exogenous gene operably linked to a mammalian promoter;
- and
- (ii) cell culture media comprising one or both of (a) D-mannosamine and (b) N-acetyl-D-mannosamine.
A nucleic acid comprising a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises
- (i) an exogenous gene under the control of a mammalian-active promoter and
- (ii) one or both of (a) a nucleic acid sequence encoding a mammalian siayltransferase and (b) a nucleic acid sequence encoding a mammalian galactosyltransferase.
A nucleic acid comprising
- (i) a genome of a non-mammalian DNA virus, wherein the genome of the virus comprises an exogenous gene under the control of a mammalian-active promoter and
- (ii) one or both of (a) a nucleic acid sequence encoding CD59 or a homolog thereof and (b) a nucleic acid sequence encoding decay accelerating factor or a homolog thereof.
A non-mammalian DNA virus wherein
- the genome of the virus comprises an exogenous gene operably linked to a mammalian-active promoter;
- and
- a coat protein of the non-mammalian DNA virus comprises a mannose core region linked to a carbohydrate moiety selected from the group consisting of N-acetyl glucosamine, galactose, and neuraminic acid.
Dependencies are inferred from the claim text.