Multicolor Fluorescence In Situ Hybridization (FISH) Approaches for Simultaneous Analysis of the Entire Human Genome

Abstract

Analysis of the organization of the human genome is vital for understanding genetic diversity, human evolution, and disease pathogenesis. A number of approaches, such as multicolor fluorescence in situ hybridization (FISH) assays, cytogenomic microarray (CMA), and next-generation sequencing (NGS) technologies, are available for simultaneous analysis of the entire human genome. Multicolor FISH-based spectral karyotyping (SKY), multiplex FISH (M-FISH), and Rx-FISH may provide rapid identification of interchromosomal and intrachromosomal rearrangements as well as the origin of unidentified extrachromosomal elements. Recent advances in molecular cytogenetics have made it possible to efficiently examine the entire human genome in a single experiment at much higher resolution and specificity using CMA and NGS technologies. Here, we present an overview of the approaches available for genome-wide analyses. © 2018 by John Wiley & Sons, Inc.

Natural language processing charts transcriptome evolution to design combination cancer therapies

“…For multiplex fluorescence in situ, hybridisation (M-FISH), 10µl of 24 colours human M-FISH paint probe mix generated at the Wellcome Trust Sanger Institute as previously described 60 was denatured at 65°C for 10 min and then applied to the denatured slides. Slide denaturation was performed by immersing slides into alkaline denaturation solution (0.5M NaOH and 1M NaCl) for 40 seconds before being rinsed with 1M Tris-HCl at pH 7.4 for 3 minutes followed by 1X PBS for 3 minutes and then dehydrated with 70%, 90% and then 100% ethanol.…”

A guide for single‐particle chromatin tracking in live cell nuclei

“…The development of fluorescence in situ hybridization (FISH) allows one to detect and locate sequence‐specific DNA/RNA in fixed cells by utilizing probes with a sequence complementary to the target sequence (Bayani & Squire, 2004; Beliveau et al, 2015). In the past few years, FISH has been extensively optimized: MerFISH, QFISH, single‐molecule fluorescence in situ hybridization (smFISH), seqFISH, CASFISH strategies improved the specificity, sensitivity, multiplexing, throughput and qualitative analysis (Deng et al, 2015; Haimovich & Gerst, 2019; Iourov, 2017; Moffitt & Zhuang, 2016; Shah et al, 2018; Xia et al, 2019; Zhang et al, 2018). Although it has advanced the field significantly, FISH suffers from inability to elucidate dynamics in the living environment.…”

“…Stable fluorescence, great multiplexing capacity, numbers of modification Fixed sample (Beliveau et al, 2015;Deng et al, 2015;Haimovich & Gerst, 2019;Iourov, 2017;Shah et al, 2018;Xia et al, 2019;Zhang et al, 2018) Repressor-operator (Lindhout et al, 2007;Lindhout et al, 2010) TALE Yes Nonintrusive (Ma et al, 2013;Thanisch et al, 2014)…”

“…Restricted to PAM sequences, local DNA unwinding/triple helix formation, potential background noise (Anton et al, 2014;Chen et al, 2013;Deng et al, 2015;Knight et al, 2015;Ma et al, 2016;Shechner et al, 2015) RNA based (Beliveau et al, 2015;Deng et al, 2015;Haimovich & Gerst, 2019;Iourov, 2017;Shah et al, 2018;Xia et al, 2019;Zhang et al Therefore, locus-specific labeling in live cells is in high demand.…”

Molecular Cytogenetics in the Era of Chromosomics and Cytogenomic Approaches

“…Cytogenetics has a low resolution of 5–10 mega base pairs, but enables a whole genomic view; it is cost-efficient and single cell oriented; i.e., it is able to pick up small mosaics. Retrospectively one can state that molecular cytogenetics was developed with the following goals: 1) to take still advantage of possibilities of banding cytogenetics, but 2) to overcome the limitation of its low resolution, and 3) to include the possibility to analyze interphase cells, too ( Zhang et al, 2018 ). Between 1969 and 1986, in situ hybridization (ISH) could exclusively be performed as a radioactive variant.…”