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DNA modification of live cell surface.

Borisenko GG, Zaitseva MA, Chuvilin AN, Pozmogova GE - Nucleic Acids Res. (2009)

Bottom Line: By using fluorescence microscopy and flow cytometry we demonstrated that our synthetic conjugates of fatty acid with oligonucleotides can be incorporated in plasma membrane and then hybridized with complementary sequences at the cell surface.All procedures can be completed within minutes and do not alter cell viability.Using this approach we tethered floating myeloid HL-60 cells to adherent A431 epitheliocytes in a sequence specific fashion.

View Article: PubMed Central - PubMed

Affiliation: Research Institute of Physical-Chemical Medicine, Russian Academy of Science, Moscow 119312, Russia. grigoryb@yahoo.com

ABSTRACT
We report a novel approach for the attachment of DNA fragments to the surface of live cells. By using fluorescence microscopy and flow cytometry we demonstrated that our synthetic conjugates of fatty acid with oligonucleotides can be incorporated in plasma membrane and then hybridized with complementary sequences at the cell surface. Method permits to control amount of immobilized DNA on the cell surface. All procedures can be completed within minutes and do not alter cell viability. Using this approach we tethered floating myeloid HL-60 cells to adherent A431 epitheliocytes in a sequence specific fashion. Thus, this method allows rapid and simple DNA multicoding of the cell surface and, therefore, opens new opportunities in manipulating with cell-cell interactions.

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Flow cytometry detection of FT18NSte integration into Jurkat cells. Cell distribution by FAM fluorescence in log scale. Curves represent populations of cells treated with 0.0. 0.05, 0.1 and 0.2 µM concentrations of FT18NSte (A) and FT18N (B) (red, black, green and blue lines, respectively). (C) Mean values of FAM fluorescence of Jurkat cells recalculated from plots of cell distribution (n = 4). Incubation conditions are same as in Figures 4A and B.
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Figure 5: Flow cytometry detection of FT18NSte integration into Jurkat cells. Cell distribution by FAM fluorescence in log scale. Curves represent populations of cells treated with 0.0. 0.05, 0.1 and 0.2 µM concentrations of FT18NSte (A) and FT18N (B) (red, black, green and blue lines, respectively). (C) Mean values of FAM fluorescence of Jurkat cells recalculated from plots of cell distribution (n = 4). Incubation conditions are same as in Figures 4A and B.

Mentions: We applied flow cytometry to quantitatively analyze incorporation of FT18NSte into cell PM. Cell distribution as measured by FAM fluorescence depended on the amount of FT18NSte added to cells, but not on the amount of FT18N (Figure 5A and B). Dependence of the fluorescence mean value on FT18NSte concentration was close to linear in the range of 0.05–0.2 µM (Figure 5C). Fluorescence intensity detected from FT18NSte-labeled cells was at least 300 times higher than from FT18N-labeled cells.Figure 5.


DNA modification of live cell surface.

Borisenko GG, Zaitseva MA, Chuvilin AN, Pozmogova GE - Nucleic Acids Res. (2009)

Flow cytometry detection of FT18NSte integration into Jurkat cells. Cell distribution by FAM fluorescence in log scale. Curves represent populations of cells treated with 0.0. 0.05, 0.1 and 0.2 µM concentrations of FT18NSte (A) and FT18N (B) (red, black, green and blue lines, respectively). (C) Mean values of FAM fluorescence of Jurkat cells recalculated from plots of cell distribution (n = 4). Incubation conditions are same as in Figures 4A and B.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651772&req=5

Figure 5: Flow cytometry detection of FT18NSte integration into Jurkat cells. Cell distribution by FAM fluorescence in log scale. Curves represent populations of cells treated with 0.0. 0.05, 0.1 and 0.2 µM concentrations of FT18NSte (A) and FT18N (B) (red, black, green and blue lines, respectively). (C) Mean values of FAM fluorescence of Jurkat cells recalculated from plots of cell distribution (n = 4). Incubation conditions are same as in Figures 4A and B.
Mentions: We applied flow cytometry to quantitatively analyze incorporation of FT18NSte into cell PM. Cell distribution as measured by FAM fluorescence depended on the amount of FT18NSte added to cells, but not on the amount of FT18N (Figure 5A and B). Dependence of the fluorescence mean value on FT18NSte concentration was close to linear in the range of 0.05–0.2 µM (Figure 5C). Fluorescence intensity detected from FT18NSte-labeled cells was at least 300 times higher than from FT18N-labeled cells.Figure 5.

Bottom Line: By using fluorescence microscopy and flow cytometry we demonstrated that our synthetic conjugates of fatty acid with oligonucleotides can be incorporated in plasma membrane and then hybridized with complementary sequences at the cell surface.All procedures can be completed within minutes and do not alter cell viability.Using this approach we tethered floating myeloid HL-60 cells to adherent A431 epitheliocytes in a sequence specific fashion.

View Article: PubMed Central - PubMed

Affiliation: Research Institute of Physical-Chemical Medicine, Russian Academy of Science, Moscow 119312, Russia. grigoryb@yahoo.com

ABSTRACT
We report a novel approach for the attachment of DNA fragments to the surface of live cells. By using fluorescence microscopy and flow cytometry we demonstrated that our synthetic conjugates of fatty acid with oligonucleotides can be incorporated in plasma membrane and then hybridized with complementary sequences at the cell surface. Method permits to control amount of immobilized DNA on the cell surface. All procedures can be completed within minutes and do not alter cell viability. Using this approach we tethered floating myeloid HL-60 cells to adherent A431 epitheliocytes in a sequence specific fashion. Thus, this method allows rapid and simple DNA multicoding of the cell surface and, therefore, opens new opportunities in manipulating with cell-cell interactions.

Show MeSH
Related in: MedlinePlus