title: Enhancing titres of therapeutic viral vectors using the transgene repression in vector production (TRiP) system.
abstract: A key challenge in the field of therapeutic viral vector/vaccine manufacturing is maximizing production. For most vector platforms, the 'benchmark' vector titres are achieved with inert reporter genes. However, expression of therapeutic transgenes can often adversely affect vector titres due to biological effects on cell metabolism and/or on the vector virion itself. Here, we exemplify the novel 'Transgene Repression In vector Production' (TRiP) system for the production of both RNA- and DNA-based viral vectors. The TRiP system utilizes a translational block of one or more transgenes by employing the bacterial tryptophan RNA-binding attenuation protein (TRAP), which binds its target RNA sequence close to the transgene initiation codon. We report enhancement of titres of lentiviral vectors expressing Cyclo-oxygenase-2 by 600-fold, and adenoviral vectors expressing the pro-apoptotic gene Bax by >150,000-fold. The TRiP system is transgene-independent and will be a particularly useful platform in the clinical development of viral vectors expressing problematic transgenes.
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: H E
authors.initials: HE
authors.lastname: Maunder
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: J
authors.initials: J
authors.lastname: Wright
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: B R
authors.initials: BR
authors.lastname: Kolli
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: C R
authors.initials: CR
authors.lastname: Vieira
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: T T
authors.initials: TT
authors.lastname: Mkandawire
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: S
authors.initials: S
authors.lastname: Tatoris
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: V
authors.initials: V
authors.lastname: Kennedy
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: S
authors.initials: S
authors.lastname: Iqball
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: G
authors.initials: G
authors.lastname: Devarajan
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: S
authors.initials: S
authors.lastname: Ellis
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: Y
authors.initials: Y
authors.lastname: Lad
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: N G
authors.initials: NG
authors.lastname: Clarkson
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: K A
authors.initials: KA
authors.lastname: Mitrophanous
authors.affiliation: Research Department, Oxford BioMedica Ltd., Windrush Court, Transport Way, Oxford OX4 6LT, UK.
authors.forename: D C
authors.initials: DC
authors.lastname: Farley
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D001412:Bacillus subtilis / Q000235:genetics; D003062:Codon; D051546:Cyclooxygenase 2 / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics; D019076:Transgenes
chemical_list: D003062:Codon; D049452:Green Fluorescent Proteins; D012313:RNA; D051546:Cyclooxygenase 2