A lmod1a mutation causes megacystis microcolon intestinal hypoperistalsis in a CRISPR/Cas9-modified zebrafish model
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In brief
In brief
Researchers used CRISPR/Cas9 to create a zebrafish model of MMIHS by deleting the lmod1a gene, successfully replicating the intestinal hypoperistalsis seen in human patients. The model showed reduced expression of key smooth muscle genes and impaired gut motility, offering a platform for testing future therapies including drug screening and gene repair approaches for this rare congenital disorder.
- CRISPR/Cas9-induced lmod1a deletion in zebrafish successfully models MMIHS with intestinal hypoperistalsis phenotype.
- Lmod1a mutation causes downregulation of smooth muscle genes (myh11, acta2) and proteins critical to intestinal motility.
- Mutant zebrafish show quantifiable peristalsis defects: fewer, slower, and shorter contractions versus wild-type controls.
- This zebrafish MMIHS model enables future drug screening and gene therapy development for this rare congenital myopathy.
- Five-base-pair deletion in lmod1a exon 1 creates premature stop codon, recapitulating human MMIHS genetic pathophysiology.
Written by the GCMD Library team from the article.
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.