Trans-splicing Into Highly Abundant Albumin Transcripts for Production of Therapeutic Proteins In Vivo

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...

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Published inMolecular therapy Vol. 17; no. 2; pp. 343 - 351
Main Authors Wang, Jun, Mansfield, S Gary, Cote, Colette A, Jiang, Ping Du, Weng, Ke, Amar, Marcelo JA, Brewer, Bryan H, Remaley, Alan T, McGarrity, Gerard J, Garcia-Blanco, Mariano A, Puttaraju, M
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.02.2009
Elsevier Limited
Nature Publishing Group
Subjects
Albumins - genetics
Amino acids
Animals
Apolipoprotein A-I - genetics
Apolipoprotein A-I - physiology
Efficiency
Exons - genetics
Female
Gene therapy
Genetic Therapy - methods
Genetic Vectors - genetics
Hemophilia
Humans
Mice
Mice, Inbred C57BL
Original
Peptides
Proteins
Reverse Transcriptase Polymerase Chain Reaction
RNA Precursors - genetics
RNA Splicing - genetics
RNA Splicing - physiology
Spliceosomes - genetics
Spliceosomes - metabolism
Trans-Splicing - genetics
Trans-Splicing - physiology
United States > US
Maryland
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Summary: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.
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Present address: Cardiovascular Research Institute, MedStar Research Institute, Washington Hospital Center, Washington, District of Columbia, USA
The first two authors contributed equally to this work.
ISSN:1525-0016
1525-0024
DOI:10.1038/mt.2008.260