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Intranuclear diffusion and hybridization state of oligonucleotides measured by fluorescence correlation spectroscopy
J C Politz1, E S Browne, D E Wolf
1Worcester Foundation for Biomedical Research, University of Massachusetts Medical Center, Worcester Foundation Campus, Shrewsbury, MA 01545, USA. politz@sci.wfbr.edu
Abstract:
Fluorescein-labeled oligodeoxynucleotides (oligos) were introduced into cultured rat myoblasts, and their molecular movements inside the nucleus were studied by fluorescence correlation spectroscopy (FCS) and fluorescence recovery after photobleaching (FRAP). FCS revealed that a large fraction of both intranuclear oligo(dT) (43%) and oligo(dA) (77%) moves rapidly with a diffusion coefficient of 4 x 10(-7) cm2/s. Interestingly, this rate of intranuclear oligo movement is similar to their diffusion rates measured in aqueous solution. In addition, we detected a large fraction (45%) of the intranuclear oligo(dT), but not oligo(dA), diffusing at slower rates (
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