An induction gene trap for identifying a homeoprotein-regulated locus

被引:27
作者
Mainguy, G
Montesinos, ML
Lesaffre, B
Zevnik, B
Karasawa, M
Kothary, R
Wurst, W
Prochiantz, A
Volovitch, M
机构
[1] Max Planck Inst Psychiat, D-80804 Munich, Germany
[2] Ecole Normale Super, CNRS, UMR 8542, F-75230 Paris 05, France
[3] GSF Forschungszentrum Umwelt & Gesundheit, D-85764 Oberschleissheim, Germany
[4] Ottawa Gen Hosp, Res Inst, Ctr Mol Med, Ottawa, ON K1H 8L6, Canada
[5] Univ Paris 07, UFR Biol, F-75005 Paris, France
关键词
gene trap; nervous system; electroporation; homeoprotein; BPAG1;
D O I
10.1038/77312
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
An important issue in developmental biology is the identification of homeoprotein target genes. We have developed a strategy based on the internalization and nuclear addressing of exogenous homeodomains, using an engrailed homeodomain (EnHD) to screen an embryonic stem (ES) cell gene trap library. Eight integrated gene trap loci responded to EnHD. One is within the bullous pemphigoid antigen 1 (BPAG1) locus, in a region that interrupts two neural isoforms. By combining in vivo electroporation with organotypic cultures, we show that an already identified BPAG1 enhancer/promoter is differentially regulated by homeoproteins Hoxc-8 and Engrailed in the embryonic spinal cord and mesencephalon. This strategy can therefore be used for identifying and mutating homeoprotein targets. Because homeodomain third helices can internalize proteins, peptides, phosphopeptides, and antisense oligonucleotides, this strategy should be applicable to other intracellular targets for characterizing genetic networks involved in a large number of physiopathological states.
引用
收藏
页码:746 / 749
页数:4
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