Amyotrophic Lateral Sclerosis study with OTHER, RNA-Seq + 1 more data on dbGaP (3 participants)

phs002440 — Parallel CRISPR Editing of Familial ALS Mutations

<p>Induced pluripotent stem cells (iPSCs) provide a modeling system for human disease within the critical context of the human genome. CRISPR editing of iPSCs allows the construction of isogenic pairs where the cells have nearly identical genomes except for the desired edited change. Here, we used CRISPR editing on a healthy control line to generate three new isogenic iPSC lines that each contains a different familial amyotrophic lateral sclerosis (fALS) mutation (<a href="https://www.ncbi.nlm.nih.gov/gene/23435">TARDBP</a>[G298S/+], <a href="https://www.ncbi.nlm.nih.gov/gene/6647">SOD1</a>[G85R/+], and <a href="https://www.ncbi.nlm.nih.gov/gene/5216">PFN1</a>[G118V/+]). As controls for each mutation, we retained iPSC lines that went through the editing process but did not have the desired heterozygous mutation. We have also performed editing in a patient cell line with a hexanucleotide repeat in <a href="https://www.ncbi.nlm.nih.gov/gene/?term=203228">C9ORF72</a>. This edit mutates a non-canonical start codon upstream of the hexanucleotide repeat. These edited lines have been used in three different studies:</p><p></p><div>(1) In “Human sensorimotor organoids derived from healthy and amyotrophic lateral sclerosis stem cells form neuromuscular junctions” (<a href="https://pubmed.ncbi.nlm.nih.gov/34362895/">PMID: 34362895</a>), we performed exome sequencing to confirm the absence of off-target CRISPR events. We also performed single-cell sequencing on sensorimotor organoids from a second healthy control iPSC line, which is deposited alongside the exome sequencing data.</div><br><div>(2) In “iPSC motor neurons, but not other derived cell types, capture gene expression changes in postmortem sporadic ALS motor neurons (<a href="https://pubmed.ncbi.nlm.nih.gov/37651231/">PMID: 37651231</a>), we differentiated these isogenic lines into motor neurons, sensory neurons, cortical neurons, and astrocytes and then performed bulk RNA-sequencing. We then assessed similarities between fALS mutations, differences between cell types, and the capacity of iPSC modeling to capture differentially expressed genes in postmortem sporadic ALS motor neurons.</div><br><div>(3) In "Blocking RAN translation without altering RNA foci rescues C9ORF72-related ALS/FTD phenotypes" (Jiang et al. 2025), we mutate a non-canonical start codon upstream of a hexanucleotide repeat in <a href="https://www.ncbi.nlm.nih.gov/gene/?term=203228">C9ORF72</a>. This mitigates several phenotypes related to neurodegeneration.</div><br><p></p>