Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes

Brian J Beliveau; Alistair N Boettiger; Maier S Avendaño; Ralf Jungmann; Ruth B McCole; Eric F Joyce; Caroline Kim-Kiselak; Frédéric Bantignies; Chamith Y Fonseka; Jelena Erceg; Mohammed A Hannan; Hien G Hoang; David Colognori; Jeannie T Lee; William M Shih; Peng Yin; Xiaowei Zhuang; Chao-ting Wu

doi:10.1038/ncomms8147

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Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes

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Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes

Brian J Beliveau, Alistair N Boettiger, Maier S Avendaño, Ralf Jungmann, Ruth B McCole, Eric F Joyce, Caroline Kim-Kiselak, Frédéric Bantignies, Chamith Y Fonseka, Jelena Erceg, …

Nature communications, Vol.6(1), p.7147

12/05/2015

DOI: https://doi.org/10.1038/ncomms8147

PMID: 25962338

Résumé

Animals

Cell Line

Chromosome Painting - methods

Chromosomes - genetics

Drosophila

Gene Library

Haplotypes

In Situ Hybridization, Fluorescence - methods

Oligonucleotide Array Sequence Analysis - methods

Oligonucleotide Probes - metabolism

Staining and Labeling

Fluorescence in situ hybridization (FISH) is a powerful single-cell technique for studying nuclear structure and organization. Here we report two advances in FISH-based imaging. We first describe the in situ visualization of single-copy regions of the genome using two single-molecule super-resolution methodologies. We then introduce a robust and reliable system that harnesses single-nucleotide polymorphisms (SNPs) to visually distinguish the maternal and paternal homologous chromosomes in mammalian and insect systems. Both of these new technologies are enabled by renewable, bioinformatically designed, oligonucleotide-based Oligopaint probes, which we augment with a strategy that uses secondary oligonucleotides (oligos) to produce and enhance fluorescent signals. These advances should substantially expand the capability to query parent-of-origin-specific chromosome positioning and gene expression on a cell-by-cell basis.

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Détails

Titre: Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes
Créateurs - sans rôle: Brian J Beliveau - Harvard Medical School
Alistair N Boettiger - 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA Howard Hughes Medical Institute, Cambridge, Massachusetts 02138, USA
Maier S Avendaño - 1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA
Ralf Jungmann - 1] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA
Ruth B McCole - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Eric F Joyce - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Caroline Kim-Kiselak - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Frédéric Bantignies - Institut de Génétique Humaine
Chamith Y Fonseka - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Jelena Erceg - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Mohammed A Hannan - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Hien G Hoang - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
David Colognori - 1] Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA Howard Hughes Medical Institute, Boston, Massachusetts 02114, USA Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA
Jeannie T Lee - 1] Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA Howard Hughes Medical Institute, Boston, Massachusetts 02114, USA Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA
William M Shih - Harvard University
Peng Yin - Harvard University
Xiaowei Zhuang - 1] Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA Howard Hughes Medical Institute, Cambridge, Massachusetts 02138, USA Department of Physics, Harvard University, Cambridge, Massachusetts 02138, USA
Chao-ting Wu - Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
Détails de publication: Nature communications, Vol.6(1), p.7147
Note de subvention: R01 GM061936 / NIGMS NIH HHS DP2 OD007292 / NIH HHS Howard Hughes Medical Institute 5DP1GM106412 / NCCDPHP CDC HHS R01-GM090278 / NIGMS NIH HHS DP2 OD004641 / NIH HHS R01 GM090278 / NIGMS NIH HHS R01GM61936 / NIGMS NIH HHS 1DP2OD004641 / NIH HHS DP1 GM106412 / NIGMS NIH HHS
Identifiants: 99159188309311
Unité académique: Institut de Génétique Humaine - IGH
Langue: English
Type de ressource: Journal article