PROJECT SUMMARY
There is a critical need to develop high-throughput scalable assays to identify biological mechanisms underlying
risk genes in neurodevelopmental and neuropsychiatric disorders (NPD). In this proposal, we aim to leverage
the unique advantages of two scalable systems – human induced pluripotent stem cells (hiPSCs) and zebrafish
– to perform parallel functional assays of NPD genes in vitro and in vivo, and to pilot the development of
innovative spatial multi-omics technologies applicable across systems. We propose to establish an Assay and
Data Generation Center (ADGC) as part of the SSPsyGene Consortium that capitalizes on the unique and
complementary expertise of our labs in large-scale hiPSC CRISPR screens (Brennand), high-throughput
zebrafish screens (Hoffman), and cutting-edge multi-omics tool development (Fan). Our goal is to gain novel
insights into the convergent and divergent mechanisms by which diverse NPD gene loss of function
affects neurodevelopment at the molecular, cellular, structural, circuit, and behavioral levels. We propose
to screen 250 NPD genes using a tiered strategy in hiPSCs and zebrafish by conducting pooled and arrayed
transcriptomic and phenotypic screens in hiPSCs-derived neurons and glia (Aim 1), CRISPR screens in
zebrafish to assess the effects of gene loss of function on whole-brain structure, activity, and basic behaviors
(Aim 2), and spatial transcriptomic and multi-omic CRISPR screens to investigate the transcriptional effects of
NPD gene disruption in both systems (Aim 3). We will advance the field by identifying biologically relevant
phenotypes resulting from NPD gene loss of function across multiple scales, informing gene prioritization
schema, and establishing new spatial multi-omics platforms for the functional analysis of NPD genes. These
studies will generate an unprecedented resource of matched molecular, cellular, structural, circuit, and
behavioral data in hiPSCs and zebrafish, which will be provided for open distribution to the broader community
to yield new insights into NPD.
