Related Experiment Video
Updated: Feb 22, 2026
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
Stable cell lines of human SH-SY5Y uniformly expressing wild-type or mutant-type FERM domain containing 7 gene
Jiali Pu1, Yanfang Mao1, Lingjia Xu1
1Department of Neurology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China.
Abstract:
It has been reported that FERM domain containing 7 (FRMD7) may cause X-linked idiopathic congenital nystagmus (ICN). A total of >40 mutations of the FRMD7 gene have been identified, however their pathogenic role remains unclear. In the present study, enhanced green fluorescent protein-tagged wild-type (WT) and mutant (MT) FRMD7 (c. C781>G) were expressed in stably expressing human neuroblastoma SH-SY5Y cells following viral transfection and antibiotic selection. Uniform expression of the FRMD7 fusion proteins was confirmed via fluorescence microscopy and western blotting. The expression profiles of neuron-specific proteins and Rho guanine triphosphatases (GTPases) differed significantly between the wild-type and mutant cell lines. Levels of Mtap2, NF-M, nestin, GAP43 and Rac1 mRNA were significantly increased in MT-FRMD7 cells compared with controls (P<0.01). However, the expression of Rac1 protein did not differ significantly among the two cell lines. Taken together, the results of the current study suggest that MT-FRMD7 influences the expression of neuron-specific genes and Rho GTPases, which may be involved in the pathogenesis of ICN. The FRMD7 stable expression cell line may facilitate future studies investigating the role of this protein in neuronal development.
Insights
Mutant FERM domain containing 7 (FRMD7) protein influences neuron-specific gene expression and Rho GTPases, potentially contributing to X-linked idiopathic congenital nystagmus (ICN) pathogenesis. This finding aids future ICN research.
Area of Science:
- Genetics
- Neuroscience
- Cell Biology
Background:
- X-linked idiopathic congenital nystagmus (ICN) is associated with the FRMD7 gene.
- Over 40 FRMD7 mutations are known, but their pathogenic roles are unclear.
- Understanding FRMD7's function is crucial for ICN research.
Purpose of the Study:
- To investigate the pathogenic role of a specific FRMD7 mutation (c. C781>G).
- To analyze the effects of mutant FRMD7 on neuronal gene and protein expression.
- To establish a cellular model for studying FRMD7 in ICN.
Main Methods:
- Expressed wild-type (WT) and mutant (MT) FRMD7 fused to GFP in SH-SY5Y cells via viral transfection.
- Confirmed uniform protein expression using fluorescence microscopy and western blotting.
- Compared expression profiles of neuron-specific proteins and Rho GTPases between WT and MT cell lines.
Main Results:
- Significant differences in neuron-specific protein and Rho GTPase expression were observed between WT and MT-FRMD7 cells.
- mRNA levels of Mtap2, NF-M, nestin, GAP43, and Rac1 were significantly increased in MT-FRMD7 cells (P<0.01).
- Rac1 protein levels did not significantly differ between the cell lines.
Conclusions:
- Mutant FRMD7 impacts the expression of neuron-specific genes and Rho GTPases.
- These alterations may play a role in the pathogenesis of ICN.
- The established FRMD7 stable cell line provides a valuable tool for future research into ICN and neuronal development.
