A CRISPR/Cas9 zebrafish lamin A/C mutant model of muscular laminopathy

Abstract

Background: Lamin A/C gene (LMNA) mutations frequently cause cardiac and/or skeletal muscle diseases called striated muscle laminopathies. We created a zebrafish muscular laminopathy model using CRISPR/Cas9 technology to target the zebrafish lmna gene.Results: Heterozygous and homozygous lmna mutants present skeletal muscle damage at 1 day post-fertilization (dpf), and mobility impairment at 4 to 7 dpf. Cardiac structure and function analyses between 1 and 7 dpf show mild and transient defects in the lmna mutants compared to wild type (WT). Quantitative RT-PCR analysis of genes implicated in striated muscle laminopathies show a decrease in jun and nfκb2 expression in 7 dpf homozygous lmna mutants compared to WT. Homozygous lmna mutants have a 1.26-fold protein increase in activated Erk 1/2, kinases associated with striated muscle laminopathies, compared to WT at 7 dpf. Activated Protein Kinase C alpha (Pkc α), a kinase that interacts with lamin A/C and Erk 1/2, is also upregulated in 7 dpf homozygous lmna mutants compared to WT.Conclusions: This study presents an animal model of skeletal muscle laminopathy where heterozygous and homozygous lmna mutants exhibit prominent skeletal muscle abnormalities during the first week of development.Furthermore, this is the first animal model that potentially implicates Pkc α in muscular laminopathies.

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],…”

“…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 ].…”

Zebrafish Models for Skeletal Muscle Senescence: Lessons from Cell Cultures and Rodent Models

“…For example, the lmna gene, whose mutation is known to accelerate aging in rodents [ 60 ], is also present in zebrafish. According to a recently published report, the deletion of five base pairs (5bpΔ) in the second exon of the lmna gene by CRISPR/Cas9 genome editing resulted in skeletal muscle damage and impaired swimming [ 61 ]. The authors used this as a model of laminopathy, such as Emery–Dreifuss muscular dystrophy; however, it also has potential as a model of skeletal muscle senescence.…”

“…However, with the aforementioned rise of CRISPR/Cas9 technology, the number of possible models of skeletal muscle senescence has been increasing in recent years. For example, in addition to the lmna crispant described above [ 61 ], tp53 crispants have overall reduced activity and shorter migration distances [ 65 ]; Tp53 is well known for its tumor suppressor activity. Therefore, senescence is reported to be accelerated in Tp53 −/m mice [ 64 ], and the knockout of tp53 in zebrafish is predicted to partially affect the skeletal muscle aging pathway.…”

Caenorhabditis elegans models for striated muscle disorders caused by missense variants of human LMNA

“…Vertebrate models have been developed to study the effects of several laminopathic variants. Zebrafish containing a 5-bp deletion that is predicted to create an early stop codon in one or both copies of lmna have skeletal muscle defects, reduced movement, and aberrant expression of genes that are also dysregulated in laminopathy patients [ 17 ]. Lmna -/- mice lose nuclear envelope integrity, have delayed postnatal growth, and experience a rapid onset of muscular dystrophy, while Lmna +/- mice develop adult-onset DCM [ 18 ].…”