The ultimate goal of this SBIR project is to develop a novel proteomic system that will enable the real- time expression profiling of proteins in live murine embryonic stem (mES) cells and in differentiated cells derived there from. The first step involves the establishment of a library of fluorescent trap mES cell lines, each of which will harbor a single fluorescently-tagged protein. The fluorescence in each cell of a particular clonal line will serve as a reporter for the expression and subcellular localization of the tagged protein in that cell line. Such a library of clonal fluorescently-tagged lines can be subjected to quantitative fluorescence microscopy in multiwell live-cell arrays, to enable the simultaneous real-time expression profiling of multiple proteins in live cells under various culture conditions. In Phase I we demonstrated the feasibility of establishing a large and diverse protein-trap library using our approach. This proposed Phase II effort will be focused generating a library of at 1500 unique fluorescent protein-trap mES cell lines. Individual cell lines from such a library of fluorescently protein-trap mES lines can also be used as versatile protein-specific research tools as they can be manipulated in culture into different tissue-types or used for the generation of transgenic mice when desired. Many also these cell lines can also be used to design novel powerful cell-based assays. Therefore this proposed Phase II project will result in the generation of a versatile and diverse proteomic research resource in mES cells.

Public Health Relevance

The ultimate aim of this SBIR project is to develop a system that will enable researchers to monitor the location and levels of hundreds of proteins simultaneously in living cells. Such a system will be useful in researching a wide variety of biological processes, including understanding the molecular basis of a number of human diseases. Further, such a system can be applied to the identification therapeutic targets and the creation of powerful drug-screening platforms to identify potential therapeutic agents that may cure these diseases.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Small Business Innovation Research Grants (SBIR) - Phase II (R44)
Project #
5R44RR024325-04
Application #
8144268
Study Section
Special Emphasis Panel (ZRG1-IMST-E (15))
Program Officer
Sheeley, Douglas
Project Start
2010-09-16
Project End
2014-07-31
Budget Start
2011-08-01
Budget End
2012-07-31
Support Year
4
Fiscal Year
2011
Total Cost
$274,952
Indirect Cost
Name
Powerhouse Proteomic Systems, LLC
Department
Type
DUNS #
602491495
City
Zionsville
State
IN
Country
United States
Zip Code
46077