Grace Tierney

Major: Integrative Neuroscience

Bio: Senior at Fordham University, scheduled to graduate in May of 2023 with a Bachelor of Science in Cell and Molecular Neuroscience.

Title of Research: Differentiating NGN2 Neurons from Neural Progenitor Cells

Faculty Mentor: Dr. Jason Morris, Department of Natural Sciences

Abstract: Many SCN2A variants have been identified as potential risk factors for Alzheimer’s disease, making the gene one of interest for researchers studying the disease. SCN2A mutations have been previously identified as correlates for schizophrenia, autism, and epilepsy, but their contributions to Alzheimer’s disease remains unknown. The following study aims to differentiate Neurogenin-2 (NGN2) human neurons with downregulated SCN2A expression from neural progenitor stem cell lines ((NPCs). The protocol describes a lentiviral infection that alters the RNA of the NPCs to repress SCN2A through CRISPR inhibition, which delivers a guide RNA to direct a dCas9-KRAB fusion protein to the gene of interest.  Following their successful differentiation, the study validates reduced gene expression and neuronal differentiation through immunostaining and a real-time qPCR test. The results of this study demonstrated reduced SCN2A expression in the experimental cell lines compared to the control cell lines, as expected, and paves the way for future functionality studies on SCN2A.