Understanding RNA Processing Changes Due to FUS in Amyotrophic Lateral Sclerosis (ALS) Patient Cells

Overview

Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease that leads to motor neuron degradation, and in many cases, paralysis and eventual death. Fused in sarcoma (FUS) mutations have been specific to familial cases of ALS and have been noted to interfere with RNA (ribonucleic acid) processes within the cell. In this project, the main focus was to find significant RNA expression changes between the wild type and mutant FUS cells from ALS patients. This project was performed by taking the fibroblasts from control and mutant FUS-ALS patients, which were then converted into neurons through protein and gene expression changes. Then, immunofluorescence was used to visualize the mislocalization of the FUS protein in fibroblasts and to confirm the presence of certain neuronal markers in the neurons to ensure proper conversion. Through the use of RASL-sequencing, two exons within the neuronal RNA, GOLPH3L exon 4 and BBX exon 14, were misregulated in almost all of the converted mutant neurons. By performing RT-PCR, the results clearly demonstrated that the mutant FUS cells have a higher inclusion of GOLPH3L exon 4 and BBX exon 14. This shows that there is an RNA splicing misregulation between mutant FUS and control FUS RNA. This project gives a conclusive answer to the hypothesis, suggesting that there is a link between FUS mutations and RNA alterations. The future applications based on this project are immense as the RNA splicing readouts can be used as a method of testing for specific drugs in order to reverse mutant FUS effects and alleviate disease symptoms.

Competition history

• AJAS 2018 Category not listed

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