Topokaryotyping demonstrates single cell variability and stress dependent variations in nuclear envelope associated domains.

ABSTRACT: Analysis of large-scale interphase genome positioning with reference to a nuclear landmark has recently been studied using sequencing-based single cell approaches. However, these approaches are dependent upon technically challenging, time consuming and costly high throughput sequencing technologies, requiring specialized bioinformatics tools and expertise. Here, we propose a novel, affordable and robust microscopy-based single cell approach, termed Topokaryotyping, to analyze and reconstruct the interphase positioning of genomic loci relative to a given nuclear landmark, detectable as banding pattern on mitotic chromosomes. This is accomplished by proximity-dependent histone labeling, where biotin ligase BirA fused to nuclear envelope marker Emerin was coexpressed together with Biotin Acce

SUBMITTER: Jurisic A 

PROVIDER: S-EPMC6294560 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature