Trans-splicing into highly abundant albumin transcripts for production of therapeutic proteins in vivo

Mol Ther. 2009 Feb;17(2):343-51. doi: 10.1038/mt.2008.260. Epub 2008 Dec 9.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Albumins / genetics*
  • Animals
  • Apolipoprotein A-I / genetics
  • Apolipoprotein A-I / physiology
  • Exons / genetics
  • Female
  • Genetic Therapy / methods
  • Genetic Vectors / genetics
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • RNA Precursors / genetics
  • RNA Splicing / genetics
  • RNA Splicing / physiology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spliceosomes / genetics
  • Spliceosomes / metabolism
  • Trans-Splicing / genetics*
  • Trans-Splicing / physiology

Substances

  • Albumins
  • Apolipoprotein A-I
  • RNA Precursors