Deepa Manwani1, Mariann Galdass, James J Bieker. 1. Department of Molecular, Cell and Developmental Biology, Mount Sinai School of Medicine, New York, NY 10029, USA. deepa.manwani@mssn.edu
Abstract
OBJECTIVE: Targeted regulation of beta-like globin genes was studied using designer zinc finger transcription factors containing the DNA binding domain of the red cell specific transcription factor erythroid Kruppel-like factor (EKLF) fused to repression domains. METHODS: Globin gene expression was analyzed after introduction of the modified transcription factors into cell lines, embryonic stem cells and transgenic mice. RESULTS: As would be predicted, when introduced transiently into cells these transcription factors were effective in repressing the adult beta-globin promoter CACCC element, which is the natural target for EKLF. In murine erythroleukemia cells repression of the adult beta-globin gene was accompanied by a reactivation of the endogenous embryonic betaH1-globin gene. Studies in differentiated embryonic stem cells and transgenic mice confirmed the reactivation of embryonic gene expression during development. CONCLUSION: Our studies support a competition model for beta-globin gene expression and underscore the importance of EKLF in the embryonic/fetal-to-adult globin switch. They also demonstrate the feasibility of designer zinc finger transcription factors in the study of transcriptional control mechanisms at the beta-globin locus and as potential gene therapy agents for sickle cell disease and related hemoglobinopathies.
OBJECTIVE: Targeted regulation of beta-like globin genes was studied using designer zinc finger transcription factors containing the DNA binding domain of the red cell specific transcription factor erythroid Kruppel-like factor (EKLF) fused to repression domains. METHODS: Globin gene expression was analyzed after introduction of the modified transcription factors into cell lines, embryonic stem cells and transgenic mice. RESULTS: As would be predicted, when introduced transiently into cells these transcription factors were effective in repressing the adult beta-globin promoter CACCC element, which is the natural target for EKLF. In murineerythroleukemia cells repression of the adult beta-globin gene was accompanied by a reactivation of the endogenous embryonic betaH1-globin gene. Studies in differentiated embryonic stem cells and transgenic mice confirmed the reactivation of embryonic gene expression during development. CONCLUSION: Our studies support a competition model for beta-globin gene expression and underscore the importance of EKLF in the embryonic/fetal-to-adult globin switch. They also demonstrate the feasibility of designer zinc finger transcription factors in the study of transcriptional control mechanisms at the beta-globin locus and as potential gene therapy agents for sickle cell disease and related hemoglobinopathies.
Authors: Roy Drissen; Robert-Jan Palstra; Nynke Gillemans; Erik Splinter; Frank Grosveld; Sjaak Philipsen; Wouter de Laat Journal: Genes Dev Date: 2004-10-15 Impact factor: 11.361
Authors: M C Rosatelli; L Oggiano; G Battista Leoni; T Tuveri; A Di Tucci; M T Scalas; F Dore; P Pistidda; A Massa; M Longinotti Journal: Blood Date: 1989-02 Impact factor: 22.113
Authors: Y Jouvenot; V Ginjala; L Zhang; P-Q Liu; M Oshimura; A P Feinberg; A P Wolffe; Rolf Ohlsson; P D Gregory Journal: Gene Ther Date: 2003-03 Impact factor: 5.250
Authors: J M Gonzalez-Redondo; T A Stoming; K D Lanclos; Y C Gu; A Kutlar; F Kutlar; T Nakatsuji; B Deng; I S Han; V C McKie Journal: Blood Date: 1988-09 Impact factor: 22.113
Authors: Miroslawa Siatecka; Kenneth E Sahr; Sabra G Andersen; Mihaly Mezei; James J Bieker; Luanne L Peters Journal: Proc Natl Acad Sci U S A Date: 2010-08-09 Impact factor: 11.205
Authors: Antanas Planutis; Li Xue; Cecelia D Trainor; Mohan Dangeti; Kevin Gillinder; Miroslawa Siatecka; Danitza Nebor; Luanne L Peters; Andrew C Perkins; James J Bieker Journal: Development Date: 2017-02-01 Impact factor: 6.868
Authors: Susan P Perrine; Rishikesh Mankidy; Michael S Boosalis; James J Bieker; Douglas V Faller Journal: Eur J Haematol Date: 2009-02-05 Impact factor: 2.997