Generation of an NCS1 gene knockout human induced pluripotent stem cell line using CRISPR/Cas9

Summary: CRISPR-Cas9 technology combined with human induced pluripotent stem cells enables precise disease modeling in pluripotent and specialized cells. This article presents an optimized CRISPR-Cas9 pipeline called ASSURED (affordable, successful, specific, user-friendly, rapid, efficient, and deliverable) for generating gene-modified single-cell-derived knockout or single-nucleotide-polymorphism-modified knockin hiPSC clones. The article describes the steps for analyzing targeted genomic sequences, designing guide RNAs and homology repair templates, as well as the CRISPR-Cas9 delivery workflow, efficiency evaluation, automated cell isolation, and clone screening.

Summary: Chemotherapy-induced peripheral neuropathy (CIPN) is a common adverse effect with no predictive biomarkers or preventive treatments available. Using sensory neurons derived from induced pluripotent stem cells as an experimental model for CIPN, researchers found differential responses to neurotoxic drugs between cell lines from different healthy donors, showcasing potential for studying CIPN pathogenesis and neuroprotective treatments.