Kruppel-like factors (KLFs) are transcription factors that regulate developmental programs. In mice without KLF2, embryonic expression of gamma-y- and beta-H-1 globin mRNA is significantly reduced. The goal is to identify novel genes involved in primitive erythropoiesis and those regulated by KLF2. The first specific aim is to generate an expression profile of mouse primitive erythropoiesis. This involves isolating erythroid and epithelial cells by laser capture microdissection from E9.5 yolk sacs. RNA from both cell types will be hybridized to Affymetrix GeneChip(R) 430A 2.0 arrays for microarray analyses. Supervised learning methods will be utilized to identify erythroid-specific genes.
In specific aim 2, differentially expressed genes will be identified in wild-type and KLF2-/- erythroid cells with the LCM/microarray strategy. Hierarchical clustering and supervised learning methods will be employed to identify differences in gene expression due to the loss of KLF2.
For specific aim 3, differential expression of erythroid genes between wild-type and KLF2-/- embryos will be verified with quantitative RT-PCR. This work will provide novel approaches to treat individuals with beta-thalassemia and sickle cell anemia.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31DK072548-02
Application #
7475672
Study Section
Special Emphasis Panel (ZRG1-GGG-G (29))
Program Officer
Agodoa, Lawrence Y
Project Start
2006-06-01
Project End
2009-05-31
Budget Start
2007-06-01
Budget End
2008-05-31
Support Year
2
Fiscal Year
2007
Total Cost
$39,594
Indirect Cost
Name
Virginia Commonwealth University
Department
Genetics
Type
Schools of Medicine
DUNS #
105300446
City
Richmond
State
VA
Country
United States
Zip Code
23298
Pang, Christopher J; Lemsaddek, Wafaa; Alhashem, Yousef N et al. (2012) Kruppel-like factor 1 (KLF1), KLF2, and Myc control a regulatory network essential for embryonic erythropoiesis. Mol Cell Biol 32:2628-44
Redmond, Latasha C; Dumur, Catherine I; Archer, Kellie J et al. (2011) Kruppel-like factor 2 regulated gene expression in mouse embryonic yolk sac erythroid cells. Blood Cells Mol Dis 47:1-11
Redmond, Latasha C; Dumur, Catherine I; Archer, Kellie J et al. (2008) Identification of erythroid-enriched gene expression in the mouse embryonic yolk sac using microdissected cells. Dev Dyn 237:436-46