DOI: 10.1101/508150Dec 31, 2018Paper

Live cell monitoring for factors affecting genome variation

BioRxiv : the Preprint Server for Biology
Yuntao XiaDennis E Discher

Abstract

Cancer cells and pluripotent stem cells frequently exhibit gains or losses of entire chromosomes and chromosome segments, and the typical terminal analyses of genomes suggest this aneuploidy is ongoing and particularly variable in solid tumors. Here, we quantify aneuploidy-inducing perturbations by live cell fluorescence monitoring for changes in chromosome-5 in a lung cancer line and in normal diploid iPS cells. Inhibition of the spindle assembly checkpoint (SAC) and knockdown of DNA repair factors cause chromosome mis-segregation to increase several-fold above a low baseline level, and both perturbations also generate several-fold more rare fluorescent-null cells. Loss of chromosome-5 is confirmed by single cell karyotyping, SNP arrays on stable isolated clones, and downregulated expression of genes on chromosome-5 in single cell transcriptomics. The iPS cells also show loss of fluorescence in infrequent cells after SAC inhibition and upon growth as teratomas in mice. Viability, selection, and altered expression can thus be tracked to reveal molecular mechanisms in aneuploidy.

Related Concepts

Aneuploidy
Chromosomes
Chromosomes, Human, Pair 5
Genes
Genome
Laboratory mice
Teratoma
Cell Line, Tumor
Fluorescence Spectroscopy
Solid Tumour

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