Abstract

The long term objective of this research plan is to define the molecular mechanisms which control globin gene expression in humans. Once these mechanisms are defined, new therapies for treatment of the many human hemoglobinopathies may be devised.
The specific aim of this proposal is to define the nucleotide sequences which control the tissue specificity and developmental specificity of human globin gene expression. In order to identify these regulatory sequences, cloned human globin genes will be microinjected into the nuclei of fertilized mouse eggs and the expression of these genes examined in erythroid and non-erythroid tissues throughout development. If the injected genes contain all of the sequences required for correct tissue specific and developmental specific expression, globin mRNA and protein will be detected only in erythroid tissues at the proper developmental stage. Once animals which correctly express human genes are obtained, modified genes which are either constructed in vitro or isolated from patients with precisely defined hemoglobinopathies will be microinjected. Modifications which alter or abolish correct tissue specific or developmental specific expression will identify important regulatory sequences.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL035559-01
Application #
3349553
Study Section
Molecular Biology Study Section (MBY)
Project Start
1985-12-01
Project End
1988-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Alabama Birmingham
Department
Type
School of Medicine & Dentistry
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294

  Publications

Levasseur, Dana N; Ryan, Thomas M; Reilly, Michael P et al. (2004) A recombinant human hemoglobin with anti-sickling properties greater than fetal hemoglobin. J Biol Chem 279:27518-24
Zhou, Dewang; Ren, Jin-Xiang; Ryan, Thomas M et al. (2004) Rapid tagging of endogenous mouse genes by recombineering and ES cell complementation of tetraploid blastocysts. Nucleic Acids Res 32:e128
Masuoka, Howard C; Townes, Tim M (2002) Targeted disruption of the activating transcription factor 4 gene results in severe fetal anemia in mice. Blood 99:736-45
Chen, W Y; Bailey, E C; McCune, S L et al. (1997) Reactivation of silenced, virally transduced genes by inhibitors of histone deacetylase. Proc Natl Acad Sci U S A 94:5798-803
Farmer, S C; Sun, C W; Winnier, G E et al. (1997) The bZIP transcription factor LCR-F1 is essential for mesoderm formation in mouse development. Genes Dev 11:786-98
Pawlik, K M; Sun, C W; Higgins, N P et al. (1995) End joining of genomic DNA and transgene DNA in fertilized mouse eggs. Gene 165:173-81
Ciavatta, D J; Ryan, T M; Farmer, S C et al. (1995) Mouse model of human beta zero thalassemia: targeted deletion of the mouse beta maj- and beta min-globin genes in embryonic stem cells. Proc Natl Acad Sci U S A 92:9259-63
Donze, D; Townes, T M; Bieker, J J (1995) Role of erythroid Kruppel-like factor in human gamma- to beta-globin gene switching. J Biol Chem 270:1955-9
Pawlik, K M; Townes, T M (1995) Autonomous, erythroid-specific DNase I hypersensitive site formed by human beta-globin locus control region (LCR) 5' HS 2 in transgenic mice. Dev Biol 169:728-32
McCune, S L; Reilly, M P; Chomo, M J et al. (1994) Recombinant human hemoglobins designed for gene therapy of sickle cell disease. Proc Natl Acad Sci U S A 91:9852-6

Showing the most recent 10 out of 20 publications