Revisiting a selection of target genes for the hematopoietic transcription factor c-Myb using chromatin immunoprecipitation and c-Myb knockdown

被引:27
作者
Berge, Tone
Matre, Vilborg
Brendeford, Elen M.
Saether, Thomas
Luescher, Bernhard
Gabrielsen, Odd Stokke [1 ]
机构
[1] Univ Oslo, Dept Mol Biosci, N-0316 Oslo, Norway
[2] Klinkum RWTH, Inst Biochem, Dept Biochem & Mol Biol, D-52074 Aachen, Germany
关键词
c-Myb; ChIP; siRNA; target gene;
D O I
10.1016/j.bcmd.2007.05.007
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The transcription factor c-Myb is an important regulator of hematopoiesis required for proper development of most blood cell lineages in vertebrates. An increasing number of target genes for c-Myb are being published, although with little or no overlap between the lists of genes reported. This raises the question of which criteria a bona fide c-Myb-target gene should satisfy. In the present paper, we have analyzed a set of previously reported target genes using chromatin immunoprecipitation (ChIP) and siRNA-mediated knockdown. Among the seven well-studied c-Myb target genes that we analyzed by ChIP, only ADA, c-MYC and MAT2A seemed to be occupied by c-Myb under our experimental settings in the Myb-positive cell lines Jurkat and HL60. After siRNA-mediated knockdown of c-Myb expression, the expression levels of two out of three ChIP positive Myb target genes, ADA and c-MYC, were strongly affected. These results clearly demonstrate the importance of combining different methods for target gene validation and suggest that a combination of ChIP and c-Myb knockdown may represent a powerful approach to identify a core collection of c-Myb target genes. (C) 2007 Elsevier Inc. All rights reserved.
引用
收藏
页码:278 / 286
页数:9
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