Modeling Sarcoglycanopathy in Danio rerio

Area of Science:

• Genetics
• Zebrafish Models
• Muscular Dystrophy Research

Background:

• Sarcoglycanopathies, a group of rare muscular dystrophies, lead to severe disability and complications like respiratory and cardiac issues.
• Current treatments are lacking, highlighting the need for effective strategies and reliable animal models for drug development.

Purpose of the Study:

• To generate and characterize zebrafish models for sarcoglycanopathies using CRISPR/Cas9 gene editing.
• To investigate disease progression and pathogenic mechanisms in these novel models.
• To establish a platform for phenotype-based drug screening.

Main Methods:

• CRISPR/Cas9 genome editing to create delta-sarcoglycan and beta-sarcoglycan knockout zebrafish.
• Phenotypic analysis of knockout zebrafish at different life stages.
• Utilizing a viscous swimming medium to accelerate disease onset detection.
• Investigating protein degradation pathways, specifically ERAD, in a delta-sarcoglycan missense mutant.

Main Results:

• Generated stable delta-sarcoglycan and beta-sarcoglycan knockout zebrafish lines.
• Observed progressive myopathic phenotypes in knockout zebrafish, worsening with age.
• Successfully used a viscous swimming assay to anticipate disease onset in larvae.
• Demonstrated that a delta-SG missense mutant is subject to ERAD, indicating misfolding and premature degradation.

Conclusions:

• Zebrafish are a valuable model for studying sarcoglycanopathies, applicable for both knockout and potential knock-in strategies.
• The developed zebrafish lines provide crucial tools for understanding disease pathogenesis.
• These models will accelerate drug screening and the development of therapeutic interventions for sarcoglycanopathies.