A 191-kb Genomic Fragment Containing the Human Alpha-globin Locus Can Rescue Alpha-thalassemic Mice

Overview

Journal Mamm Genome

Specialty Genetics

Date 2005 Nov 15

PMID 16284800

Citations 5

Authors

Keith Al-Hasani

Jim Vadolas

Anja S Knaupp

Hady Wardan

Lucille Voullaire

Robert Williamson

Panayiotis A Ioannou

Affiliations

Soon will be listed here.

Abstract

A 191-kb human bacterial artificial chromosome (BAC) containing the human alpha-globin genomic locus was used to generate transgenic mice that express, exclusively, human alpha-globin ((hu)alpha-globin). Expression of (hu)alpha-globin reaches a level of 36% of that of endogenous mouse alpha-globin ((mu)alpha-globin) on a heterozygous mouse alpha-thalassemia background ((mu)alpha-globin knockout, (mu)alpha(+/-)). Hemizygous transgenic mice carrying the (hu)alpha-globin locus on a heterozygous knockout background ((hu)alpha(+/0), (mu)alpha(++/--)) demonstrated complementation of most hematologic parameters. By crossing (hu)alpha(+/0), (mu)alpha(++/--) mice, we were able to generate mice entirely dependent on (hu)alpha-globin synthesis. Breeding and fluorescent in situ hybridization studies demonstrate that only mice homozygous for the transgene were able to rescue embryonic lethal homozygous (mu)alpha-globin knockout embryos ((mu)alpha(--/--)). Adult rescued mice produce hemoglobin at levels similar to wild-type mice, with partial red cell complementation based on mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and red cell distribution width (RDW) measurements. Significant erythrocythemia above wild-type levels seems to be the main compensatory mechanism for the normalization of the hemoglobin levels in the rescued animals. Our studies demonstrate that the (hu)alpha-globin locus in the 191-kb transgene contains all the necessary elements for the regulated expression of (hu)alpha-globin in transgenic mice. This animal model should be valuable for studying the mechanisms regulating (hu)alpha-globin production and for development of therapeutic strategies for beta-thalassemia based on downregulation of alpha-globin expression.

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References

Mortlock D, Guenther C, Kingsley D . A general approach for identifying distant regulatory elements applied to the Gdf6 gene. Genome Res. 2003; 13(9):2069-81. PMC: 403689. DOI: 10.1101/gr.1306003. View

Vadolas J, Wardan H, Orford M, Voullaire L, Zaibak F, Williamson R . Development of sensitive fluorescent assays for embryonic and fetal hemoglobin inducers using the human beta -globin locus in erythropoietic cells. Blood. 2002; 100(12):4209-16. DOI: 10.1182/blood-2001-12-0365. View

Vadolas J, Wardan H, Bosmans M, Zaibak F, Jamsai D, Voullaire L . Transgene copy number-dependent rescue of murine beta-globin knockout mice carrying a 183 kb human beta-globin BAC genomic fragment. Biochim Biophys Acta. 2005; 1728(3):150-62. DOI: 10.1016/j.bbaexp.2005.02.008. View

Jamsai D, Zaibak F, Khongnium W, Vadolas J, Voullaire L, Fowler K . A humanized mouse model for a common beta0-thalassemia mutation. Genomics. 2005; 85(4):453-61. DOI: 10.1016/j.ygeno.2004.11.016. View

Sarsero J, Li L, Holloway T, Voullaire L, Gazeas S, Fowler K . Human BAC-mediated rescue of the Friedreich ataxia knockout mutation in transgenic mice. Mamm Genome. 2004; 15(5):370-82. DOI: 10.1007/s00335-004-3019-3. View

Vadolas J, Wardan H, Orford M, Williamson R, Ioannou P . Cellular genomic reporter assays for screening and evaluation of inducers of fetal hemoglobin. Hum Mol Genet. 2003; 13(2):223-33. DOI: 10.1093/hmg/ddh023. View

Feng D, Liu D, Huang Y, Wu L, Li T, Wu M . The expression of human alpha -like globin genes in transgenic mice mediated by bacterial artificial chromosome. Proc Natl Acad Sci U S A. 2001; 98(26):15073-7. PMC: 64985. DOI: 10.1073/pnas.251550598. View

Hatton C, Wilkie A, DRYSDALE H, Wood W, Vickers M, Sharpe J . Alpha-thalassemia caused by a large (62 kb) deletion upstream of the human alpha globin gene cluster. Blood. 1990; 76(1):221-7. View

Higgs D, Vickers M, Wilkie A, Pretorius I, Jarman A, Weatherall D . A review of the molecular genetics of the human alpha-globin gene cluster. Blood. 1989; 73(5):1081-104. View

10.

Higgs D, Sharpe J, Wood W . Understanding alpha globin gene expression: a step towards effective gene therapy. Semin Hematol. 1998; 35(2):93-104. View

11.

Schrier S . Pathophysiology of thalassemia. Curr Opin Hematol. 2002; 9(2):123-6. DOI: 10.1097/00062752-200203000-00007. View

12.

Paszty C, Mohandas N, Stevens M, Loring J, Liebhaber S, Brion C . Lethal alpha-thalassaemia created by gene targeting in mice and its genetic rescue. Nat Genet. 1995; 11(1):33-9. DOI: 10.1038/ng0995-33. View

13.

Gourdon G, Sharpe J, Wells D, Wood W, Higgs D . Analysis of a 70 kb segment of DNA containing the human zeta and alpha-globin genes linked to their regulatory element (HS-40) in transgenic mice. Nucleic Acids Res. 1994; 22(20):4139-47. PMC: 331901. DOI: 10.1093/nar/22.20.4139. View

14.

Al-Hasani K, Vadolas J, Voullaire L, Williamson R, Ioannou P . Complementation of alpha-thalassaemia in alpha-globin knockout mice with a 191 kb transgene containing the human alpha-globin locus. Transgenic Res. 2004; 13(3):235-43. DOI: 10.1023/b:trag.0000034627.36174.06. View

15.

Rund D, RACHMILEWITZ E . Pathophysiology of alpha- and beta-thalassemia: therapeutic implications. Semin Hematol. 2001; 38(4):343-9. DOI: 10.1016/s0037-1963(01)90028-9. View

16.

Leder A, Daugherty C, Whitney B, Leder P . Mouse zeta- and alpha-globin genes: embryonic survival, alpha-thalassemia, and genetic background effects. Blood. 1997; 90(3):1275-82. View