POXVIRUS HOST RANGE PROTEIN K3 AS A POSITIVE SELECTION MARKER FOR GENERATION OF RECOMBINANT POXVIRUSES, A THERAPEUTIC TARGET FOR POXVIRUS INFECTION AND A THERAPEUTIC AGENT FOR PKR RELATED DISEASES

摘要

Described herein is a novel method for generation of recombinant poxviruses using an E3 and K3 double deletion mutant virus as the parental virus for generation of recombinant viruses. Following allowing for crossing over between the parental virus and an insertion cassette including an orthopox K3 peptide and the gene of interest, recombinant viruses are selected by infecting a host cell line permissive for the orthopox K3 peptide but not for the E3 and K3 double mutant parental virus. It is also demonstrated that a specific small molecule inhibitor of NEDD8 activating enzyme, MLN4924, can completely block poxvirus K3 family protein mediated PKR degradation and virus replication.