PUBLICATION
A lmod1a mutation causes megacystis microcolon intestinal hypoperistalsis in a CRISPR/Cas9-modified zebrafish model
- Authors
- Kalim, A.S., Nagata, K., Toriigahara, Y., Shirai, T., Kirino, K., Xiu-Ying, Z., Kondo, T., Kawakubo, N., Miyata, J., Matsuura, T., Tajiri, T.
- ID
- ZDB-PUB-240815-10
- Date
- 2024
- Source
- Pediatric surgery international 40: 225225 (Journal)
- Registered Authors
- Keywords
- Animal model, CRISPR/Cas9, Lmod1a, MMIHS, Zebrafish
- MeSH Terms
-
- Abnormalities, Multiple/genetics
- Animals
- CRISPR-Cas Systems*
- Colon*/abnormalities
- Disease Models, Animal*
- Intestinal Pseudo-Obstruction*/genetics
- Muscle Proteins/genetics
- Mutation
- Urinary Bladder/abnormalities
- Zebrafish*/genetics
- Zebrafish Proteins/genetics
- PubMed
- 39143337 Full text @ Pediatr. Surg. Int.
Citation
Kalim, A.S., Nagata, K., Toriigahara, Y., Shirai, T., Kirino, K., Xiu-Ying, Z., Kondo, T., Kawakubo, N., Miyata, J., Matsuura, T., Tajiri, T. (2024) A lmod1a mutation causes megacystis microcolon intestinal hypoperistalsis in a CRISPR/Cas9-modified zebrafish model. Pediatric surgery international. 40:225225.
Abstract
Purpose Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is defined as a congenital visceral myopathy with genetic mutations. However, the etiology and pathophysiology are not fully understood. We aimed to generate a gene leiomodin-1a (lmod1a) modification technique to establish a zebrafish model of MMIHS.
Methods We targeted lmod1a in zebrafish using CRISPR/Cas9. After confirming the genotype, we measured the expression levels of the target gene and protein associated with MMIHS. A gut transit assay and spatiotemporal mapping were conducted to analyze the intestinal function.
Results Genetic confirmation showed a 5-base-pair deletion in exon 1 of lmod1a, which caused a premature stop codon. We observed significant mRNA downregulation of lmod1a, myh11, myod1, and acta2 and the protein expression of Lmod1 and Acta2 in the mutant group. A functional analysis of the lmod1a mutant zebrafish showed that its intestinal peristalsis was fewer, slower, and shorter in comparison to the wild type.
Conclusion This study showed that targeted deletion of lmod1a in zebrafish resulted in depletion of MMIHS-related genes and proteins, resulting in intestinal hypoperistalsis. This model may have the potential to be utilized in future therapeutic approaches, such as drug discovery screening and gene repair therapy for MMIHS.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping