|Start Date / Time||September 21, 2022 at 9:00 AM|
|End Date / Time||September 21, 2022 at 10:00 AM|
|Presenter Name||Mirazul Islam|
|Presentation Title||“Developmental and temporal recording of whole mouse at single cell resolution”|
|Status||This meeting has already occurred|
Students and Postdocs
CRISPR-Cas9-based technologies have been developed to address a wide range of biological questions, including molecular recording of embryonic development, genetic screening, and tracking tumor evolution at the single-cell resolution. We developed a single-cell lineage tracking platform called Native sgRNA Capture and sequencing (NSC-seq) to leverage CRISPR technology to decipher early lineage branching point of mouse gastrulation. We combined CRISPR-induced mutations with spontaneously accumulating mitochondrial variants to enable highly efficient lineage-tracking of single cells. Applying this platform in a CRISPR-based transgenic mouse line (MARC1) allows us to reconstruct both embryonic and adult developmental lineages at single-cell resolution. In addition, we performed clonal analysis of intestinal tumors in the barcoded ApcMin/+ mouse model and found that there are more than 2 independently arising clones within a tumor, supporting the oligoclonality of intestinal tumor development. Finally, we apply NSC-seq to assess a comprehensive gene expression phenotype for individual gene knockout at the single-cell level using existing whole-genome CRISPR knockout screening plasmid libraries. Overall, NSC-seq enables in vivo developmental and temporal recording and single-cell CRISPR screening at single cell resolution.