Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 109, Issue 52, Pages 21301-21306Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1213818110
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Funding
- National Institutes of Health (NIH)/National Institute of General Medical Sciences [1R01GM085169]
- Broad Institute Scientific Planning and Allocation of Resources Committee (SPARC) Award
- Harvard Medical School
- NSF
- NIH/National Cancer Institute Ruth L. Kirschstein National Research Service Award
- NIH Centers of Excellence in Genomic Science
- NIH [5R42GM097003, 1R43GM093579]
- [1S10RR028832-01]
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A host of observations demonstrating the relationship between nuclear architecture and processes such as gene expression have led to a number of new technologies for interrogating chromosome positioning. Whereas some of these technologies reconstruct intermolecular interactions, others have enhanced our ability to visualize chromosomes in situ. Here, we describe an oligonucleotide- and PCR-based strategy for fluorescence in situ hybridization (FISH) and a bioinformatic platform that enables this technology to be extended to any organism whose genome has been sequenced. The oligonucleotide probes are renewable, highly efficient, and able to robustly label chromosomes in cell culture, fixed tissues, and metaphase spreads. Our method gives researchers precise control over the sequences they target and allows for single and multicolor imaging of regions ranging from tens of kilobases to megabases with the same basic protocol. We anticipate this technology will lead to an enhanced ability to visualize interphase and metaphase chromosomes.
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