This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Construction of an inducible plasmid: For regulated expression of enhanced green fluorescent protein (EGFP), lentiviral vectors with doxycycline inducible promoter system based on hybrid repressor tetR-VP16 or tetR-KRAB can be used. In absence of doxycycline, the hybrid repressor will bind specifically to tetO and suppresses the nearby promoter controlling EGFP expression. Conversely, in the presence of doxycycline, the repressor will be sequestered away from tetO, thus permitting expression of EGFP. tetR: tetracyclin repressor; VP16: HSV VP16 transactivator domain; KRAB: KRAB domain of human Kox1; tetO: tet operator sequenceOnce the constructs are ready the hESC will be transfected and checked for GFP expression ad injectedin to E13.5 mouse embryonic kidney and organ cultured for 7 days. The kidenys will then be subjected to immno-histochemical analysis.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Exploratory Grants (P20)
Project #
5P20RR016475-08
Application #
7720198
Study Section
Special Emphasis Panel (ZRR1-RI-7 (02))
Project Start
2008-05-01
Project End
2009-04-30
Budget Start
2008-05-01
Budget End
2009-04-30
Support Year
8
Fiscal Year
2008
Total Cost
$32,204
Indirect Cost
Name
University of Kansas
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
016060860
City
Kansas City
State
KS
Country
United States
Zip Code
66160
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