Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes

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.

[1]  T. Xie,et al.  Identification of Genes That Promote or Antagonize Somatic Homolog Pairing Using a High-Throughput FISH–Based Screen , 2012, PLoS genetics.

[2]  Hideki Tanizawa,et al.  Unravelling global genome organization by 3C-seq. , 2012, Seminars in cell & developmental biology.

[3]  J Vrolijk,et al.  New strategy for multi-colour fluorescence in situ hybridisation: COBRA: COmbined Binary RAtio labelling , 1999, European Journal of Human Genetics.

[4]  Hao Yan,et al.  Self-assembled combinatorial encoding nanoarrays for multiplexed biosensing. , 2007, Nano letters.

[5]  Carolyn J. Brown,et al.  A gene from the region of the human X inactivation centre is expressed exclusively from the inactive X chromosome , 1991, Nature.

[6]  J. Gall,et al.  Molecular hybridization of radioactive DNA to the DNA of cytological preparations. , 1969, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[8]  W. D. Laat,et al.  A Decade of 3c Technologies: Insights into Nuclear Organization References , 2022 .

[9]  M. Ploeg,et al.  Use of whole cosmid cloned genomic sequences for chromosomal localization by non-radioactive in situ hybridization , 1987, Human Genetics.

[10]  F. Simmel,et al.  Single-molecule kinetics and super-resolution microscopy by fluorescence imaging of transient binding on DNA origami. , 2010, Nano letters.

[11]  Michael Wigler,et al.  PROBER: oligonucleotide FISH probe design software , 2006, Bioinform..

[12]  Jay Shendure,et al.  Multiplex amplification of large sets of human exons , 2007, Nature Methods.

[13]  D. Ledbetter,et al.  Multicolor Spectral Karyotyping of Human Chromosomes , 1996, Science.

[14]  Carol J. Bult,et al.  Folding and organization of a contiguous chromosome region according to the gene distribution pattern in primary genomic sequence , 2006, The Journal of cell biology.

[15]  Matthew J. Rodesch,et al.  Fluorescence in situ hybridization with high-complexity repeat-free oligonucleotide probes generated by massively parallel synthesis , 2011, Chromosome Research.

[16]  T. Cremer,et al.  Chromosome territories. , 2010, Cold Spring Harbor perspectives in biology.

[17]  F S Fay,et al.  Visualization of single RNA transcripts in situ. , 1998, Science.

[18]  A. Tsalenko,et al.  Visualization of Fine-Scale Genomic Structure by Oligonucleotide-Based High-Resolution FISH , 2010, Cytogenetic and Genome Research.

[19]  J. Kolberg,et al.  Single-copy Gene Detection Using Branched DNA (bDNA) In Situ Hybridization , 2001, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[20]  William C Earnshaw,et al.  Super-resolution fluorescence microscopy as a tool to study the nanoscale organization of chromosomes. , 2011, Current opinion in chemical biology.

[21]  Scott A. Rifkin,et al.  Imaging individual mRNA molecules using multiple singly labeled probes , 2008, Nature Methods.

[22]  D. Ward,et al.  Karyotyping human chromosomes by combinatorial multi-fluor FISH , 1996, Nature Genetics.

[23]  H. Dalbøge,et al.  Detection of proopiomelanocortin mRNA by in situ hybridization, using a biotinylated oligodeoxynucleotide probe and avidin-alkaline phosphatase histochemistry , 2004, Histochemistry.

[24]  Robert H. Singer,et al.  Fluorescence in situ hybridization: past, present and future , 2003, Journal of Cell Science.

[25]  A. Oudenaarden,et al.  Validating transcripts with probes and imaging technology , 2011, Nature Methods.

[26]  A. Silahtaroglu,et al.  FISHing with locked nucleic acids (LNA): evaluation of different LNA/DNA mixmers. , 2003, Molecular and cellular probes.

[27]  Y. Dufrêne Atomic force microscopy of membrane proteins separating two aqueous compartments , 2006, Nature Methods.

[28]  D. Ward,et al.  Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries , 1988, Human Genetics.

[29]  H. Tanke,et al.  Heterogeneity in telomere length of human chromosomes. , 1996, Human molecular genetics.

[30]  R. Hawley,et al.  c(3)G encodes a Drosophila synaptonemal complex protein. , 2001, Genes & development.

[31]  C. Wright Structural comparison of the two distinct sugar binding sites in wheat germ agglutinin isolectin II. , 1984, Journal of molecular biology.

[32]  C. O'keefe,et al.  Oligonucleotide probes for alpha satellite DNA variants can distinguish homologous chromosomes by FISH. , 1996, Human molecular genetics.

[33]  Penghua Zhang,et al.  Discovery of natural nicking endonucleases Nb.BsrDI and Nb.BtsI and engineering of top-strand nicking variants from BsrDI and BtsI , 2007, Nucleic acids research.

[34]  X. Zhuang,et al.  Breaking the Diffraction Barrier: Super-Resolution Imaging of Cells , 2010, Cell.

[35]  Alexander van Oudenaarden,et al.  A versatile genome-scale PCR-based pipeline for high-definition DNA FISH , 2013, Nature Methods.

[36]  J. Maguire,et al.  Solution Hybrid Selection with Ultra-long Oligonucleotides for Massively Parallel Targeted Sequencing , 2009, Nature Biotechnology.

[37]  M. Macek,et al.  The Use of Peptide Nucleic Acids for In Situ Identification of Human Chromosomes , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[38]  J. Birchler,et al.  Single-Gene Detection and Karyotyping Using Small-Target Fluorescence in Situ Hybridization on Maize Somatic Chromosomes , 2007, Genetics.

[39]  R. Jaenisch,et al.  A 450 kb Transgene Displays Properties of the Mammalian X-Inactivation Center , 1996, Cell.

[40]  Chiara Lanzuolo,et al.  Polycomb response elements mediate the formation of chromosome higher-order structures in the bithorax complex , 2007, Nature Cell Biology.

[41]  Andrew D Griffiths,et al.  Amplification of complex gene libraries by emulsion PCR , 2006, Nature Methods.

[42]  Christoph Cremer,et al.  COMBO-FISH Enables High Precision Localization Microscopy as a Prerequisite for Nanostructure Analysis of Genome Loci , 2010, International journal of molecular sciences.

[43]  Wendy A Bickmore,et al.  Chromatin organization in the mammalian nucleus. , 2005, International review of cytology.

[44]  E. Joyce,et al.  Cytological analysis of meiosis in fixed Drosophila ovaries. , 2009, Methods in molecular biology.

[45]  K. Johansen,et al.  Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling , 2010, Journal of visualized experiments : JoVE.

[46]  I. Amit,et al.  Comprehensive mapping of long range interactions reveals folding principles of the human genome , 2011 .

[47]  E. Volpi,et al.  FISH glossary: an overview of the fluorescence in situ hybridization technique. , 2008, BioTechniques.

[48]  Michael Zuker,et al.  UNAFold: software for nucleic acid folding and hybridization. , 2008, Methods in molecular biology.

[49]  D. Agard,et al.  Perturbation of Nuclear Architecture by Long-Distance Chromosome Interactions , 1996, Cell.

[50]  J. Dekker,et al.  Capturing Chromosome Conformation , 2002, Science.

[51]  E. Heard,et al.  Combined immunofluorescence, RNA fluorescent in situ hybridization, and DNA fluorescent in situ hybridization to study chromatin changes, transcriptional activity, nuclear organization, and X-chromosome inactivation. , 2008, Methods in molecular biology.

[52]  Andrew Lawler,et al.  Treaty Draft Raises Scientific Hackles , 1996, Science.

[53]  Dan Luo,et al.  Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes , 2005, Nature Biotechnology.

[54]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[55]  M. Zuker,et al.  OligoArray 2.0: design of oligonucleotide probes for DNA microarrays using a thermodynamic approach. , 2003, Nucleic acids research.

[56]  J. Wiegant,et al.  A new method for fluorescence microscopical localization of specific DNA sequences by in situ hybridization of fluorochromelabelled RNA. , 1980, Experimental cell research.

[57]  M. Moreno,et al.  Applications of peptide nucleic acids (PNAs) and locked nucleic acids (LNAs) in biosensor development , 2012, Analytical and Bioanalytical Chemistry.