Fluorescence imaging of chromosomal DNA using click chemistry.
Chromosome visualization is essential for chromosome analysis and genetic diagnostics. Here, we developed a click chemistry approach for multicolor imaging of chromosomal DNA instead of the traditional dye method. We first demonstrated that the commercially available reagents allow for the multicolor staining of chromosomes. We then prepared two pro-fluorophore moieties that served as light-up reporters to stain chromosomal DNA based on click reaction and visualized the clear chromosomes in multicolor. We applied this strategy in fluorescence in situ hybridization (FISH) and identified, with high sensitivity and specificity, telomere DNA at the end of the chromosome. We further extended this approach to observe several basic stages of cell division. We found that the click reaction enables direct visualization of the chromosome behavior in cell division. These results suggest that the technique can be broadly used for imaging chromosomes and may serve as a new approach for chromosome analysis and genetic diagnostics.
Division of Chemistry, Department of Medical Sciences, Faculty of Medicine, University of Miyazaki, Japan.
Takumi
T
Ishizuka
Division of Chemistry, Department of Medical Sciences, Faculty of Medicine, University of Miyazaki, Japan.
Hong Shan
HS
Liu
Research Center for Advanced Science and Technology, The University of Tokyo, Japan.
Kenichiro
K
Ito
Division of Chemistry, Department of Medical Sciences, Faculty of Medicine, University of Miyazaki, Japan.
Yan
Y
Xu
D002455:Cell Division / Q000235:genetics; D057930:Click Chemistry / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D003952:Diagnostic Imaging / Q000379:methods; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015203:Reproducibility of Results; D016615:Telomere / Q000235:genetics*
D005456:Fluorescent Dyes; D004247:DNA