This research project aims to create isogenic model of human induced pluripotent stem cells (iPSCs) with NRXN1α gene deletion as carried by patients with Autism Spectrum Disorder (ASD), using the CRISPR/Cas9 gene-editing technology, which won the 2020 Nobel Prize. NRXN1α is one of the major rare genetic factors, contributing to the development of ASD. The research seeks to minimize the effects of genetic background and heterogeneity by generating NRXN1α deletions from multiple iPSC lines of healthy controls, so that isogenic NRXN1α+/- iPSCs can be studied systematically by comparison with healthy parental iPSC lines. This study aims to create human isogenic iPSCs with NRXN1α deletion that can be used to study physiological and pathological mechanisms of ASD. The objectives of the study include designing and synthesizing guide RNAs (gRNAs) to target the NRXN1α gene, electroporation of iPSCs with gRNAs and Cas9, screening and validating successful NRXN1α deletion clones by genomic PCR and DNA sequencing. The research intends to elucidate the functional consequence of the NRXN1α deletion in human brain, particularly in the fundamental process of neuronal activities: excitation and inhibition balance. It will clarify the role of NRXN1α isoform deletion in the mechanisms behind NRXN1α deletions in individuals with ASD. The findings of the study can contribute to a deeper understanding of the genetic and cellular mechanisms underlying ASD and may potentially lead to the development of new therapeutic interventions for this neurodevelopmental disorder.
