Modeling Sarcoglycanopathy in Danio rerio

Abstract

Sarcoglycanopathies, also known as limb girdle muscular dystrophy 3-6, are rare muscular dystrophies characterized, although heterogeneous, by high disability, with patients often wheelchair-bound by late adolescence and frequently developing respiratory and cardiac problems. These diseases are currently incurable, emphasizing the importance of effective treatment strategies and the necessity of animal models for drug screening and therapeutic verification. Using the CRISPR/Cas9 genome editing technique, we generated and characterized δ-sarcoglycan and β-sarcoglycan knockout zebrafish lines, which presented a progressive disease phenotype that worsened from a mild larval stage to distinct myopathic features in adulthood. By subjecting the knockout larvae to a viscous swimming medium, we were able to anticipate disease onset. The δ-SG knockout line was further exploited to demonstrate that a δ-SG missense mutant is a substrate for endoplasmic reticulum-associated degradation (ERAD), indicating premature degradation due to protein folding defects. In conclusion, our study underscores the utility of zebrafish in modeling sarcoglycanopathies through either gene knockout or future knock-in techniques. These novel zebrafish lines will not only enhance our understanding of the disease’s pathogenic mechanisms, but will also serve as powerful tools for phenotype-based drug screening, ultimately contributing to the development of a cure for sarcoglycanopathies.

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

“…In creating disease models, known mutations in genes that cause particular genetic syndromes in humans are obtained, such as muscle laminopathy [ 31 ], Charlevoix-Saguenay spastic ataxia [ 32 ], Marfan syndrome [ 33 ], Sanfilippo syndrome [ 34 ], Joubert syndrome [ 35 ], Bernard-Soulier syndrome [ 36 ], Xia-Gibbs syndrome [ 37 ], Lee syndrome [ 38 ], Laron syndrome [ 39 ], Aicardi-Gutierrez syndrome [ 40 ], Finnish-type nephrotic syndrome [ 41 ], fragile cornea syndrome [ 42 ], multicentric carpotarsal osteolysis syndrome [ 43 ], Bietti crystalline dystrophy [ 44 ], sarcoglycanopathy [ 45 ], autosomal recessive microcephaly [ 46 ], sphingolipidosess [ 47 ], and mitochondrial diseases caused by polg gene mutations [ 48 ].…”