Horsman Sebastiaan, Moorhouse Michael J, de Jager Victor C L, van der Spek Peter, Grosveld Frank, Strouboulis John, Katsantoni Eleni Z
Department of Cell Biology, Erasmus Medical Center, Dr Molewaterplein 50, 3015GE Rotterdam, The Netherlands.
BMC Bioinformatics. 2006 Mar 8;7:120. doi: 10.1186/1471-2105-7-120.
In the current era of high throughput genomics a major challenge is the genome-wide identification of target genes for specific transcription factors. Chromatin immunoprecipitation (ChIP) allows the isolation of in vivo binding sites of transcription factors and provides a powerful tool for examining gene regulation. Crosslinked chromatin is immunoprecipitated with antibodies against specific transcription factors, thus enriching for sequences bound in vivo by these factors in the immunoprecipitated DNA. Cloning and sequencing the immunoprecipitated sequences allows identification of transcription factor target genes. Routinely, thousands of such sequenced clones are used in BLAST searches to map their exact location in the genome and the genes located in the vicinity. These genes represent potential targets of the transcription factor of interest. Such bioinformatics analysis is very laborious if performed manually and for this reason there is a need for developing bioinformatic tools to automate and facilitate it.
In order to facilitate this analysis we generated TF Target Mapper (Transcription Factor Target Mapper). TF Target Mapper is a BLAST search tool allowing rapid extraction of annotated information on genes around each hit. It combines sequence cleaning/filtering, pattern searching and BLAST searches with extraction of information on genes located around each BLAST hit and comparisons of the output list of genes or gene ontology IDs with user-implemented lists. We successfully applied and tested TF Target Mapper to analyse sequences bound in vivo by the transcription factor GATA-1. We show that TF Target Mapper efficiently extracted information on genes around ChIPed sequences, thus identifying known (e.g. alpha-globin and zeta-globin) and potentially novel GATA-1 gene targets.
TF Target Mapper is a very efficient BLAST search tool that allows the rapid extraction of annotated information on the genes around each hit. It can contribute to the comprehensive bioinformatic transcriptome/regulome analysis, by providing insight into the mechanisms of action of specific transcription factors, thus helping to elucidate the pathways these factors regulate.
在当前高通量基因组学时代,一个主要挑战是全基因组范围内鉴定特定转录因子的靶基因。染色质免疫沉淀(ChIP)技术能够分离转录因子在体内的结合位点,并为研究基因调控提供了一个强大的工具。交联染色质用针对特定转录因子的抗体进行免疫沉淀,从而在免疫沉淀的DNA中富集这些因子在体内结合的序列。对免疫沉淀序列进行克隆和测序,可鉴定转录因子的靶基因。通常,数千个这样的测序克隆用于BLAST搜索,以确定它们在基因组中的精确位置以及附近的基因。这些基因代表了感兴趣转录因子的潜在靶标。如果手动进行这种生物信息学分析会非常费力,因此需要开发生物信息学工具来实现自动化并简化这一过程。
为了便于进行这种分析,我们开发了TF靶标定位器(转录因子靶标定位器)。TF靶标定位器是一种BLAST搜索工具,能够快速提取每个命中周围基因的注释信息。它将序列清理/过滤、模式搜索和BLAST搜索与提取每个BLAST命中周围基因的信息以及将基因或基因本体ID的输出列表与用户自定义列表进行比较相结合。我们成功应用并测试了TF靶标定位器来分析转录因子GATA-1在体内结合的序列。我们表明,TF靶标定位器能够有效地提取ChIP序列周围基因的信息,从而鉴定出已知的(如α-珠蛋白和ζ-珠蛋白)以及潜在的新型GATA-1基因靶标。
TF靶标定位器是一种非常高效的BLAST搜索工具,能够快速提取每个命中周围基因的注释信息。它可以通过深入了解特定转录因子的作用机制,为全面的生物信息学转录组/调控组分析做出贡献,从而有助于阐明这些因子调控的途径。