Split intein-mediated protein trans-splicing to express large dystrophins

Hichem Tasfaout (), Christine L. Halbert, Timothy S. McMillen, James M. Allen, Theodore R. Reyes, Galina V. Flint, Dirk Grimm, Stephen D. Hauschka, Michael Regnier and Jeffrey S. Chamberlain ()
Additional contact information
Hichem Tasfaout: University of Washington School of Medicine
Christine L. Halbert: University of Washington School of Medicine
Timothy S. McMillen: University of Washington
James M. Allen: University of Washington School of Medicine
Theodore R. Reyes: University of Washington School of Medicine
Galina V. Flint: University of Washington
Dirk Grimm: University of Heidelberg
Stephen D. Hauschka: University of Washington School of Medicine
Michael Regnier: University of Washington School of Medicine
Jeffrey S. Chamberlain: University of Washington School of Medicine

Nature, 2024, vol. 632, issue 8023, 192-200

Abstract: Abstract Gene replacement using adeno-associated virus (AAV) vectors is a promising therapeutic approach for many diseases1,2. However, this therapeutic modality is challenged by the packaging capacity of AAVs (approximately 4.7 kilobases)3, limiting its application for disorders involving large coding sequences, such as Duchenne muscular dystrophy, with a 14 kilobase messenger RNA. Here we developed a new method for expressing large dystrophins by utilizing the protein trans-splicing mechanism mediated by split inteins. We identified several split intein pairs that efficiently join two or three fragments to generate a large midi-dystrophin or the full-length protein. We show that delivery of two or three AAVs into dystrophic mice results in robust expression of large dystrophins and significant physiological improvements compared with micro-dystrophins. Moreover, using the potent myotropic AAVMYO4, we demonstrate that low total doses (2 × 1013 viral genomes per kg) are sufficient to express large dystrophins in striated muscles body-wide with significant physiological corrections in dystrophic mice. Our data show a clear functional superiority of large dystrophins over micro-dystrophins that are being tested in clinical trials. This method could benefit many patients with Duchenne or Becker muscular dystrophy, regardless of genotype, and could be adapted to numerous other disorders caused by mutations in large genes that exceed the AAV capacity.

Date: 2024
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41586-024-07710-8 Abstract (text/html)
Access to the full text of the articles in this series is restricted.

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:nature:v:632:y:2024:i:8023:d:10.1038_s41586-024-07710-8

Ordering information: This journal article can be ordered from
https://www.nature.com/

DOI: 10.1038/s41586-024-07710-8

Access Statistics for this article

Nature is currently edited by Magdalena Skipper

More articles in Nature from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().