Identification of Gene Positioning Factors Using High-Throughput Imaging Mapping
Sigal Shachar1, Ty C Voss2, Gianluca Pegoraro2
1National Cancer Institute, NIH, Bethesda, MD 20892, USA.
Cell
|August 16, 2015
Summary
We developed HIPMap, a high-throughput imaging pipeline, to map genome organization in 3D cell nuclei. This method identified 50 new factors crucial for maintaining correct spatial genome positioning.
Area of Science:
- Cell Biology
- Genomics
- Molecular Biology
Background:
- Genomes are organized non-randomly in the 3D space of the cell nucleus.
- Understanding genome organization is key to cellular function.
Purpose of the Study:
- To develop a high-throughput method for mapping genome region locations.
- To identify cellular factors involved in spatial genome organization.
Main Methods:
- Developed HIPMap (High-precision, High-throughput, Automated Fluorescent In Situ Hybridization Imaging Pipeline).
- Utilized HIPMap for an unbiased siRNA screen in human cells.
- Mapped spatial locations of diverse genomic loci.
Main Results:
- Identified 50 cellular factors essential for proper genome positioning.
- Positioning factors include chromatin remodelers, histone modifiers, and nuclear proteins.
- Replication and chromatin re-assembly machinery components are key positioning factors.
- Cell cycle progression through replication, not mitosis, is critical for gene positioning.
Conclusions:
- HIPMap provides a large-scale method for mapping genome locations.
- A compendium of cellular factors involved in spatial genome organization was identified.
- This study advances our understanding of the molecular mechanisms underlying genome architecture.
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