March 6 @ 11:00 AM – 12:00 PM
Dr. Xin Jin
Associate Professor, Department of Neuroscience
Scripps Research
“Scalable investigation of gene function and organ-wide action with in vivo Perturb-seq”
Abstract: Human genetics have identified long lists of risk genes and loci associated with diseases, including neurodegenerative diseases and aging, and the next challenge is to identify their shared impacts and patterns of etiology for therapeutic designs. These genetic analyses are best performed in vivo where the age-dependent organismal physiology is intact. However, systematic analysis of gene function across diverse cell types in vivo is still hindered by at least two challenges: obtaining sufficient cells from live tissues and accurately identifying each cell’s perturbation in high-throughput single-cell assays. Leveraging AAV’s versatile cell type tropism and high labeling capacity, we expanded the resolution and scale of in vivo CRISPR screens: allowing phenotypic analysis at single-cell resolution across a multitude of cell types in the embryonic brain, adult brain, and peripheral nervous system. We undertook extensive tests of 86 AAV serotypes, combined with a transposon system, to substantially amplify labeling and accelerate in vivo gene delivery from weeks to days. Using this platform, we performed an in vivo genetic screen as proof-of-principle and identified pleiotropic regulatory networks of Foxg1, including their impact on Layer 6 corticothalamic neurons where it tightly controls distinct networks essential for cell fate specification and function. Notably, our platform can label >6% of cerebral cells, surpassing the current state-of-the-art efficacy at <0.1% (mediated by lentivirus), and achieve analysis of over 30,000 cells in one experiment, thus enabling massively parallel in vivo Perturb-seq. Compatible with various perturbation techniques (CRISPRa/i) and phenotypic measurements (single-cell or spatial multi-omics), our platform presents a flexible, modular approach to interrogate gene function across diverse cell types in vivo, translating gene variants to their causal functions.
Host: Ethan Hollingsworth
Seminar will be held in person only.
Developmental & Cell Biology Fall 2024 Seminar Series
Each quarter the Department of Developmental and Cell Biology hosts a weekly seminar. The purpose of these seminars is to enable experts from around the country to share their newest discoveries and ideas with our students and faculty. Seminars are held on Thursdays at 11:00 a.m., in Natural Sciences II room 4201.
For questions about this event, please contact Mayra Rubio at mrubio3@uci.edu.