Supplementary Materials

Supplementary Material for:

Single-cut genome editing restores dystrophin expression in a new mouse model of muscular dystrophy

Leonela Amoasii, Chengzu Long, Hui Li, Alex A. Mireault, John M. Shelton, Efrain Sanchez-Ortiz, John R. McAnally, Samadrita Bhattacharyya, Florian Schmidt, Dirk Grimm, Stephen D. Hauschka, Rhonda Bassel-Duby, Eric N. Olson*

*Corresponding author. Email: eric.olson{at}utsouthwestern.edu

Published 29 November 2017, Sci. Transl. Med. 9, eaan8081 (2017)
DOI: 10.1126/scitranslmed.aan8081

This PDF file includes:

  • Materials and Methods
  • Fig. S1. ΔEx50 mouse model analyses.
  • Fig. S2. Comparison of dystrophin staining in ΔEx50 and mdx mouse models at 1 month of age.
  • Fig. S3. Characterization of ΔEx50 mice at different ages.
  • Fig. S4. ESEs of exon 51.
  • Fig. S5. Validation of sgRNAs in mouse 10T1/2 and human 293T cells.
  • Fig. S6. Cas9 expression in injected muscles.
  • Fig. S7. In vivo Dmd gene editing.
  • Fig. S8. Off-target analyses for sgRNA-51.
  • Fig. S9. Off-target sequence analyses for sgRNA-51.
  • Fig. S10. Amplicon PCR deep-sequencing analyses for sgRNA-51.
  • Fig. S11. Rescue of dystrophin expression after intramuscular injection of AAV9-Cas9 and AAV9-sgRNA-51 in the ΔEx50 mouse model.
  • Fig. S12. AAV9-Cas9– and AAV9-sgRNA-51–injected muscle shows histological improvement after 3 weeks.
  • Fig. S13. Quantification of histological improvement of AAV9-Cas9– and AAV9-sgRNA-51–injected muscle from ΔEx50 mice after 3 weeks.
  • Fig. S14. Intramuscular injection of AAV9-Cas9 and AAV9-sgRNA-51 in ΔEx50 mice corrects dystrophin expression in the heart.
  • Fig. S15. AAV9-Cas9 expression after systemic delivery in mice.
  • Fig. S16. Dmd gene editing 4 weeks after systemic delivery of AAV9-Cas9 and AAV9-sgRNA-51 in mice.
  • Fig. S17. Rescue of dystrophin expression 8 weeks after systemic delivery of AAV9-Cas9 and AAV9-sgRNA-51 in ΔEx50 mice.
  • Fig. S18. Histological analysis of dystrophin correction 8 weeks after systemic delivery of AAV9-Cas9 and AAV9-sgRNA-51 in ΔEx50 mice.
  • Fig. S19. Histological improvement of ΔEx50 mice 4 weeks after systemic injection of AAV9-Cas9 and AAV9-sgRNA-51.
  • Table S1. Sequences of potential exonic off-target (OT) sites in the mouse genome.
  • Table S2. Sequences of top 45 off-target (OT) sites in the mouse genome.
  • Table S3. Primer sequences.
  • References (47, 48)

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