PUBLICATION
CRISPR/Cas9-Mediated Constitutive Loss of VCP (Valosin-Containing Protein) Impairs Proteostasis and Leads to Defective Striated Muscle Structure and Function In Vivo
- Authors
- Voisard, P., Diofano, F., Glazier, A.A., Rottbauer, W., Just, S.
- ID
- ZDB-PUB-220625-16
- Date
- 2022
- Source
- International Journal of Molecular Sciences 23(12): (Journal)
- Registered Authors
- Diofano, Federica, Just, Steffen, Rottbauer, Wolfgang
- Keywords
- CRISPR/Cas9, VCP, VCPopathies, disease modeling, protein homeostasis, zebrafish
- MeSH Terms
-
- Muscle, Skeletal/metabolism
- CRISPR-Cas Systems
- Animals
- Valosin Containing Protein/genetics
- Valosin Containing Protein/metabolism
- Proteostasis/genetics
- Cell Cycle Proteins/genetics
- Cell Cycle Proteins/metabolism
- Muscle, Striated*/metabolism
- Zebrafish/genetics
- Zebrafish/metabolism
- Adenosine Triphosphatases/genetics
- Adenosine Triphosphatases/metabolism
- Mutation
- Myositis, Inclusion Body*/genetics
- Myositis, Inclusion Body*/metabolism
- Frontotemporal Dementia*/genetics
- Frontotemporal Dementia*/metabolism
- PubMed
- 35743185 Full text @ Int. J. Mol. Sci.
Citation
Voisard, P., Diofano, F., Glazier, A.A., Rottbauer, W., Just, S. (2022) CRISPR/Cas9-Mediated Constitutive Loss of VCP (Valosin-Containing Protein) Impairs Proteostasis and Leads to Defective Striated Muscle Structure and Function In Vivo. International Journal of Molecular Sciences. 23(12).
Abstract
Valosin-containing protein (VCP) acts as a key regulator of cellular protein homeostasis by coordinating protein turnover and quality control. Mutations in VCP lead to (cardio-)myopathy and neurodegenerative diseases such as inclusion body myopathy with Paget's disease of the bone and frontotemporal dementia (IBMPFD) or amyotrophic lateral sclerosis (ALS). To date, due to embryonic lethality, no constitutive VCP knockout animal model exists. Here, we generated a constitutive CRISPR/Cas9-induced vcp knockout zebrafish model. Similar to the phenotype of vcp morphant knockdown zebrafish embryos, we found that vcp-null embryos displayed significantly impaired cardiac and skeletal muscle function. By ultrastructural analysis of skeletal muscle cells and cardiomyocytes, we observed severely disrupted myofibrillar organization and accumulation of inclusion bodies as well as mitochondrial degeneration. vcp knockout was associated with a significant accumulation of ubiquitinated proteins, suggesting impaired proteasomal function. Additionally, markers of unfolded protein response (UPR)/ER-stress and autophagy-related mTOR signaling were elevated in vcp-deficient embryos, demonstrating impaired proteostasis in VCP-null zebrafish. In conclusion, our findings demonstrate the successful generation of a stable constitutive vcp knockout zebrafish line that will enable characterization of the detailed mechanistic underpinnings of vcp loss, particularly the impact of disturbed protein homeostasis on organ development and function in vivo.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping