Researchers
at Princeton University have developed a novel technique for the simultaneous
identification of transfected cells and cell cycle analysis by flow cytometry.
Princeton is currently seeking industrial collaborators to commercialize this
technology.
The
green fluorescent protein (GFP) is currently used in assays as a marker to
detect the subpopulation of transfected cells. However, there are certain
technical limitations to the use of GFP in combination with other protocols such
as cell cycle analysis using propidium iodide (PI) and flow cytometry. For
example, in cell cycle analysis, the use of ethanol results in a loss of GFP
fluorescence as it leaches from the cell. Although prior treatment of the cells
with paraformaldehyde helps retain the GFP in the cell following ethanol
permeabilization, it precludes optimal staining of the DNA with
PI.
The
integral membrane GFP-fusion protein developed at Princeton overcomes these
limitations and produces a significant increase in the GFP signal for both the
enhanced detection of transfected cells as well as for the simultaneous
measurement of cell cycle and transfection. The GFP fusion protein is anchored
in the cell membrane and is thus retained in the cell during fixation and/or
permeabilization techniques using ethanol and may be used in combination with
other techniques to quantify cell cycle analysis (using PI) or other intra- or
extra-cellular antigens.
Patent
protection is pending.
For more
information please
contact:
William H. Gowen
Office of Technology Licensing and Intellectual
Property
Princeton University
4
New South Building
Princeton, NJ 08544-0036
(609) 258-6762
(609) 258-1159 fax
wgowen@princeton.edu