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AssigneeJanssen Pharmaceuticals, Inc.
InventorsKurt Iver Kamrud, Nathaniel Stephen Wang, Martina Felderman, Nancy C. Carrico
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The present disclosure generally relates to viral-based expression systems suitable for the production of molecule of interests in recombinant host cells. The disclosure particularly relates to nucleic acid constructs, such as expression vectors, containing a modified arterivirus genome or replicon RNA in which at least some of its original viral sequence has been deleted. Also included in the disclosure are viral-based expression vectors including one or more expression cassettes encoding heterologous polypeptides. In some embodiments, the expression cassettes are configured and positioned at defined locations on the viral genome so as to enable expression of the heterologous polypeptides in a tunable manner.
A method for producing a polypeptide of interest, comprising culturing a host cell comprising a nucleic acid, wherein the nucleic acid comprises a gene encoding the polypeptide of interest and a nucleotide sequence encoding a modified Equine arteritis virus (EAV) genome or replicon RNA, wherein the modified EAV genome or replicon RNA comprises a sequence fragment exhibiting at least 80% sequence identity to the sequence encoding open reading frame ORF7 of the wild-type sequence of EAV, and wherein the modified genome or replicon RNA is devoid of the sequence encoding ORF2a, and further wherein the polypeptide of interest is expressed.
A method for producing a polypeptide of interest in a subject, comprising administering to the subject a nucleic acid, wherein the nucleic acid comprises:
A composition comprising a nucleic acid, wherein the nucleic acid comprises a nucleotide sequence encoding a modified Equine arteritis virus (EAV) genome or replicon RNA, further wherein the nucleic acid comprises a gene encoding a polypeptide of interest, wherein the modified EAV genome or replicon RNA comprises a sequence fragment exhibiting at least 80% sequence identity to the sequence encoding open reading frame ORF7 of the wild-type sequence of EAV, and wherein the modified EAV genome or replicon RNA is devoid of the sequence encoding ORF2a;
A composition comprising a recombinant cell comprising a nucleic acid, wherein the nucleic acid comprises a nucleotide sequence encoding a modified Equine arteritis virus (EAV) genome or replicon RNA, further wherein the nucleic acid comprises a sequence encoding a gene of interest (GOI), wherein the modified EAV genome or replicon RNA comprises a sequence fragment exhibiting at least 80% sequence identity to the sequence encoding open reading frame ORF7 of the wild-type sequence of EAV and wherein the modified EAV genome or replicon RNA is devoid of the sequence encoding ORF2a, and a pharmaceutically acceptable carrier.
A method for producing a polypeptide of interest, comprising culturing a host cell comprising a nucleic acid, wherein the nucleic acid comprises a gene encoding the polypeptide of interest and a modified Equine arteritis virus (EAV) sub-genomic replicon RNA, wherein the modified EAV sub-genomic replicon RNA comprises a sequence fragment exhibiting at least 80% sequence identity to the sequence encoding open reading frame ORF7 of the wild-type sequence of EAV and further comprises a nucleotide sequence encoding at least one of (a) ORF1a and ORF1b;
Dependencies are inferred from the claim text.