title: Trans-splicing into highly abundant albumin transcripts for production of therapeutic proteins in vivo.
abstract: Spliceosome-mediated RNA trans-splicing has emerged as an exciting mode of RNA therapy. Here we describe a novel trans-splicing strategy, which targets highly abundant pre-mRNAs, to produce therapeutic proteins in vivo. First, we used a pre-trans-splicing molecule (PTM) that mediated trans-splicing of human apolipoprotein A-I (hapoA-I) into the highly abundant mouse albumin exon 1. Hydrodynamic tail vein injection of the hapoA-I PTM plasmid in mice followed by analysis of the chimeric transcripts and protein, confirmed accurate and efficient trans-splicing into albumin pre-mRNA and production of hapoA-I protein. The versatility of this approach was demonstrated by producing functional human papillomavirus type-16 E7 (HPV16-E7) single-chain antibody in C57BL/6 mice and functional factor VIII (FVIII) and phenotypic correction in hemophilia A mice. Altogether, these studies demonstrate that trans-splicing to highly abundant albumin transcripts can be used as a general platform to produce therapeutic proteins in vivo.
authors.affiliation: VIRxSYS Corporation, Gaithersburg, Maryland, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: S Gary
authors.initials: SG
authors.lastname: Mansfield
authors.forename: Colette A
authors.initials: CA
authors.lastname: Cote
authors.forename: Ping Du
authors.initials: PD
authors.lastname: Jiang
authors.forename: Ke
authors.initials: K
authors.lastname: Weng
authors.forename: Marcelo J A
authors.initials: MJ
authors.lastname: Amar
authors.forename: Bryan H
authors.initials: BH
authors.lastname: Brewer
authors.forename: Alan T
authors.initials: AT
authors.lastname: Remaley
authors.forename: Gerard J
authors.initials: GJ
authors.lastname: McGarrity
authors.forename: Mariano A
authors.initials: MA
authors.lastname: Garcia-Blanco
authors.forename: M
authors.initials: M
authors.lastname: Puttaraju
mesh_terms: D000418:Albumins / Q000235:genetics*; D000818:Animals; D016632:Apolipoprotein A-I / Q000235:genetics / Q000502:physiology; D005091:Exons / Q000235:genetics; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D012322:RNA Precursors / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017381:Spliceosomes / Q000235:genetics / Q000378:metabolism; D020040:Trans-Splicing / Q000235:genetics* / Q000502:physiology
chemical_list: D000418:Albumins; D016632:Apolipoprotein A-I; D012322:RNA Precursors