This Core is a traditional Core and has an additional role in in vivo validation of anti-sickling strategies using transgenic mice that formed a part of Project 1 in our original proposal. The Core services consist of three functions: 1) Make sure of the Transgenic and Gene Targeting Facility at the Albert Einstein College of Medicine where mice designed to achieve the goals outlined in Project 1 by RL Nagel will be generated with DNA constructs supplied by Core B under supervision of Eric Bouhassira. 2) Identify founders expressing the desired hemoglobins, characterize them, and breed them to our transgenic mice expression alpha/H/beta/S and/or the new mice created by RMCE (described in Aim 3), alpha-knockout, beta-knockout, and other transgenic and knockout lines available and backcross them onto C57BL/6 for use in Projects 1, 2, and 4, 3) Provide analytic services for other projects; protein identification, quantification (HPLC, mass spec), and red cell characterization. We will use our current well characterized models to develop methodologies for characterizing the efficacy of anti-sickling strategies Develop two new methods for evaluating hypoxia in sickle mice and those with anti-sickling globins: 1) HIF-1 alpha as a means of testing for local hypoxia. 2) Apply BOLD and flow sensitive MRI techniques to monitor for the presence of deoxyhemoglobin and improved/impaired flow in transgenic models. Use the anomalous severity in the S+S+Antilles mouse and its rescue by low levels of gamma to help design more effective anti-sickling strategies. We will take advantage of the Recombinase Mediated Cassette Exchange (RMCE) technology described in Project 2 to generate a completely new type of sickle cell mouse model to test anti-sickling globins. New sickle cell mouse models with variable levels of expression of the transgene but which are comparable in copy number and chromatin structure over the lifetime of the mouse will simplify use of these animals. Elimination of globin insufficiency, thalassemia, and comparable mice with various levels of anti-sickling globin expression will considerably simplify the interpretation of changes in pathophysiology resulting from our proposed interventions. Using this technology, we expect to be able to unambiguously determine which of the anti-sickling globins we are planning to test the best in an in vivo context.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
Project #
Application #
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Albert Einstein College of Medicine
United States
Zip Code
Dykstra, Brad; Kent, David; Bowie, Michelle et al. (2007) Long-term propagation of distinct hematopoietic differentiation programs in vivo. Cell Stem Cell 1:218-29
Bowie, Michelle B; Kent, David G; Dykstra, Brad et al. (2007) Identification of a new intrinsically timed developmental checkpoint that reprograms key hematopoietic stem cell properties. Proc Natl Acad Sci U S A 104:5878-82
Bowie, Michelle B; Kent, David G; Copley, Michael R et al. (2007) Steel factor responsiveness regulates the high self-renewal phenotype of fetal hematopoietic stem cells. Blood 109:5043-8
Dykstra, Brad; Ramunas, John; Kent, David et al. (2006) High-resolution video monitoring of hematopoietic stem cells cultured in single-cell arrays identifies new features of self-renewal. Proc Natl Acad Sci U S A 103:8185-90
Olivier, Emmanuel N; Rybicki, Anne C; Bouhassira, Eric E (2006) Differentiation of human embryonic stem cells into bipotent mesenchymal stem cells. Stem Cells 24:1914-22
Peshkovsky, Alexey; Kennan, Richard P; Nagel, Ronald L et al. (2006) Sensitivity enhancement and compensation of RF penetration artifact with planar actively detunable quadrature surface coil. Magn Reson Imaging 24:81-7
Bowie, Michelle B; McKnight, Kristen D; Kent, David G et al. (2006) Hematopoietic stem cells proliferate until after birth and show a reversible phase-specific engraftment defect. J Clin Invest 116:2808-16
Fu, Haiqing; Wang, Lixin; Lin, Chii-Mei et al. (2006) Preventing gene silencing with human replicators. Nat Biotechnol 24:572-6
Fischer, Marlene; Schmidt, Manfred; Klingenberg, Silke et al. (2006) Short-term repopulating cells with myeloid potential in human mobilized peripheral blood do not have a side population (SP) phenotype. Blood 108:2121-3
Dykxhoorn, Derek M; Schlehuber, Lisa D; London, Irving M et al. (2006) Determinants of specific RNA interference-mediated silencing of human beta-globin alleles differing by a single nucleotide polymorphism. Proc Natl Acad Sci U S A 103:5953-8

Showing the most recent 10 out of 51 publications