Identifying the gene regulatory network of neurogenin - instrument supplement

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Project Summary Schizophrenia is a highly prevalent and debilitating neurological disorder that affects between 0.3% and 0.6% of the U.S. population. There is a strong genetic component to schizophrenia, and Genome-Wide Association Studies have identified over 100 candidate loci that contribute to this disease. Among this list is the transcription factor neurogenin1. As a proneural transcription factor capable of directing neurogenesis, this gene has the potential to control the expression of multiple genes during nervous system development and function. Despite this, little is known about the identity of neurogenin’s downstream transcriptional targets, creating an imperative for further study. We propose to investigate the gene regulatory network surrounding neurogenin using the nematode C. elegans as a tractable model for these studies. We recently performed a comparative transcriptome study on C. elegans wild type and ngn-1/neurogenin mutants during embryogenesis. This identified nearly 600 genes that are under direct or indirect neurogenin transcriptional control. We hypothesize that human orthologs of neurogenin transcriptional targets have a strong possibility of being schizophrenia loci in their own right. This proposed study will use chromatin immunoprecipitation plus next-generation sequencing (ChIPseq) to identify neurogenin binding sites in the worm genome, then use these data to cross-reference against our comparative transcriptome dataset to delineate direct versus indirect neurogenin transcriptional targets. This instrument supplement will purchase a bath sonicator to facilitate chromatin preparation for these ChIPseq assays. We will validate these results using GFP reporter gene studies, with a focus on contextualizing neurogenin’s control of downstream transcriptional regulators. Finally, we will use recently published single cell RNA sequencing datasets, coupled with an unpublished ngn-1 expression lineage, to predict and validate genes that may control ngn-1’s transcription. This innovative approach will contextualize up-stream regulators as part of the neurogenin gene regulatory network. Homan orthologs of these genes may also contribute to the development of schizophrenia. This project directly addresses fundamental mechanisms of nervous system development and gene regulation. In addition, it will accomplish both broad and specific AREA program goals, including enhancing Kennesaw State University’s research environment and exposing students to high quality research through direct participation. PHS 398/2590 (Rev. 11/07) Page 1 Continuation Format Page