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Biology subjects

Shin, G.

Publications and source records attributed to Shin, G..

2 recordsLinked to original sources

Division-resolved inference of flow and trajectories in proliferating cell populations

High-throughput single-cell assays are widely used to quantify distributions of cell size, morphology, and molecular content across thousands of cells. However, such population distributions do not reveal how the measured cellular states change within individual cells over time. We introduce division-resolved inference of flow and trajectories (DRIFT), a computational framework that infers the dynamics of a measured cellular state from population distributions collected over time, without synchronizing or tracking individual cells. DRIFT solves a population-balance equation to separate state progression from the redistribution caused by cell division in proliferating populations. In simulations of growth and division perturbations, DRIFT recovered the ground-truth mean volume trajectories across simulated single-cell lineages. In live L1210 leukemia cells, DRIFT inferred perturbation-specific volume trajectories that were consistent with longitudinal single-cell measurements. Beyond cell volume, DRIFT also inferred DNA-content dynamics from fixed-cell flow cytometry in L1210 cells, consistent with independent DNA-synthesis assays. In live HeLa cells, DRIFT inferred cell area dynamics that were validated by continuous imaging. Overall, DRIFT converts endpoint measurements of cell populations into division-resolved cellular dynamics, providing a scalable strategy for high-throughput drug-response screening and mechanistic investigation.

systems biology

Assembly of Mb-size genome segments from linked read sequencing of CRISPR DNA targets

We developed a targeted sequencing method for intact high molecular weight (HMW) DNA targets as large as 0.2 Mb. This process uses HMW DNA isolated from intact cells, custom designed Cas9-guide RNA complexes to generate 0.1 - 0.2 Mb DNA targets, electrophoretic isolation of the DNA targets and sequencing with barcode linked reads. We used alignment methods as well as local assembly of the target regions to identify haplotypes and structural variants (SVs) across multi-Megabase genomic regions. To demonstrate the performance of this approach, we designed three assays that covered a 0.2 Mb region surrounding the BRCA1 gene, a set of 40 overlapping 0.2 Mb targets covering the entire 4-Mb MHC locus, and 18 well-characterized structural variants. Using the highly characterized NA12878 genome, we achieved on-target coverage of more than 50X, while overall whole genome coverage was approximately 4X. We generated haplotypes that completely covered each targeted locus, with a maximum size of 4 Mb (for the MHC region). This method detected structural variants such as deletions and inversions with determination of the exact breakpoints and genotypes. Even breakpoints inside highly homologous segmental duplications are precisely determined with our high-quality assemblies. Overall, this is a new method to sequence large DNA segments.

genomics