Chemical Engineering Seminar
Single-cell sequencing can directly reveal cell-to-cell variation at both genomic and transcriptomic levels. The digital nature and base-pair resolution of single-cell sequencing also makes it a high-resolution, high-throughput assay for studying molecular biology at the single-cell level. Here I describe a new approach ('Look-Seq') combining DNA sequencing and live-cell imaging to characterize DNA damage due to cell division errors. The Look-Seq analysis directly links genetic mutations detected by DNA sequencing to aberrant chromosomes detected by live-cell imaging by the chromosomal haplotype. Importantly, comparison of the genetic variants phased to each homologous chromosome distinguishes biological variation on a single chromosome (but not its homolog) from single-cell sequencing artifacts affecting both homologs. I discuss how DNA damage and chromosome missegregation can be measured by single-cell sequencing and how we apply the LookSeq analysis to demonstrate that an intact chromosome partitioned into abnormal structures called 'micronuclei' can lead to a catastrophic mutagenesis known as `chromothripsis', a hitherto inexplicable mutational phenomenon frequently observed in genetic disorders and cancers. Finally, I discuss applications of the Look-Seq approach in other areas of molecular biology and in cancer biology.