Spatiotemporal allele organization by allele-specific CRISPR live-cell imaging (SNP-CLING). Journal Article uri icon

Overview

abstract

  • Imaging and chromatin capture techniques have provided important insights into our understanding of nuclear organization. A limitation of these techniques is the inability to resolve allele-specific spatiotemporal properties of genomic loci in living cells. Here, we describe an allele-specific CRISPR live-cell DNA imaging technique (SNP-CLING) to provide the first comprehensive insights into allelic positioning across space and time in mouse embryonic stem cells and fibroblasts. With 3D imaging, we studied alleles on different chromosomes in relation to one another and relative to nuclear substructures such as the nucleolus. We find that alleles maintain similar positions relative to each other and the nucleolus; however, loci occupy unique positions. To monitor spatiotemporal dynamics by SNP-CLING, we performed 4D imaging and determined that alleles are either stably positioned or fluctuating during cell state transitions, such as apoptosis. SNP-CLING is a universally applicable technique that enables the dissection of allele-specific spatiotemporal genome organization in live cells.

publication date

  • February 1, 2018

Full Author List

  • Maass PG; Barutcu AR; Shechner DM; Weiner CL; MelĂ© M; Rinn JL

Other Profiles

Additional Document Info

start page

  • 176

end page

  • 184

volume

  • 25

issue

  • 2