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Cell. 2015 Aug 13;162(4):911-23. doi: 10.1016/j.cell.2015.07.035.

Identification of Gene Positioning Factors Using High-Throughput Imaging Mapping.

Author information

1
National Cancer Institute, NIH, Bethesda, MD 20892, USA.
2
High Throughput Imaging Facility (HiTIF), National Cancer Institute, NIH, Bethesda, MD 20892, USA.
3
National Cancer Institute, NIH, Bethesda, MD 20892, USA. Electronic address: mistelit@mail.nih.gov.

Abstract

Genomes are arranged non-randomly in the 3D space of the cell nucleus. Here, we have developed HIPMap, a high-precision, high-throughput, automated fluorescent in situ hybridization imaging pipeline, for mapping of the spatial location of genome regions at large scale. High-throughput imaging position mapping (HIPMap) enabled an unbiased siRNA screen for factors involved in genome organization in human cells. We identify 50 cellular factors required for proper positioning of a set of functionally diverse genomic loci. Positioning factors include chromatin remodelers, histone modifiers, and nuclear envelope and pore proteins. Components of the replication and post-replication chromatin re-assembly machinery are prominently represented among positioning factors, and timely progression of cells through replication, but not mitosis, is required for correct gene positioning. Our results establish a method for the large-scale mapping of genome locations and have led to the identification of a compendium of cellular factors involved in spatial genome organization.

PMID:
26276637
PMCID:
PMC4538709
DOI:
10.1016/j.cell.2015.07.035
[Indexed for MEDLINE]
Free PMC Article

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