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Phenotype correction of hemophilia A mice by spliceosome-mediated RNA trans-splicing

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Abstract

Conventional gene therapy of hemophilia A relies on the transfer of factor VIII (FVIII; encoded by the F8 gene) cDNA. We carried out spliceosome-mediated RNA trans-splicing (SMaRT) to repair mutant FVIII mRNA. A pre-trans-splicing molecule (PTM) corrected endogenous FVIII mRNA in F8 knockout mice with the hemophilia A phenotype, producing sufficient functional FVIII to correct the hemophilia A phenotype. This is the first description of phenotypic correction of a genetic defect by RNA repair in a knockout animal model. Our results indicate the feasibility of using SMaRT to repair RNA for the treatment of genetic diseases.

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Figure 1: Trans-splicing approach to repair of mouse F8 exon 16 mutation.
Figure 2: Trans-spliced mouse FVIII activity and antigen.
Figure 3: Quantitation of cis- and trans-spliced mouse F8 mRNA.

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Acknowledgements

We thank J. Huang and X. Zeng for technical assistance, S. Connelly and P. Shirley (GTI/Novartis) for the AdhuF8 mouse plasma and R. Sarkar and H. Kazazian for the F8 E16 knockout mice and advice. H.J.C. is a recipient of a Career Development Award from the National Hemophilia Foundation. National Institutes of Health grant RO1-68215 provided funding for this project.

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Author notes

  1. Hengjun Chao and S Gary Mansfield: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine, Mt. Sinai School of Medicine, New York, 10029, New York, USA

    Hengjun Chao & Christopher E Walsh

  2. Intronn Inc., Gaithersburg, Maryland, 20878, USA

    S Gary Mansfield, Robert C Bartel, Suja Hiriyanna, Lloyd G Mitchell & Mariano A Garcia-Blanco

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  1. Hengjun Chao
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  2. S Gary Mansfield
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  3. Robert C Bartel
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  4. Suja Hiriyanna
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  5. Lloyd G Mitchell
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  6. Mariano A Garcia-Blanco
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  7. Christopher E Walsh
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Corresponding author

Correspondence to Christopher E Walsh.

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Competing interests

S.G.M., R.C.B., S.H. and L.G.M. are employees of Intronn Inc. M.A.G.-B. is a consultant of Intronn Inc.

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Chao, H., Mansfield, S., Bartel, R. et al. Phenotype correction of hemophilia A mice by spliceosome-mediated RNA trans-splicing. Nat Med 9, 1015–1019 (2003). https://doi.org/10.1038/nm900

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