Oeder Sebastian, Mages Joerg, Flicek Paul, Lang Roland
Institute of Medical Microbiology, Immunology and Hygiene Technical University Munich, Munich, Germany.
BMC Genomics. 2007 Jun 28;8:200. doi: 10.1186/1471-2164-8-200.
The function and significance of the widespread expression of natural antisense transcripts (NATs) is largely unknown. The ability to quantitatively assess changes in NAT expression for many different transcripts in multiple samples would facilitate our understanding of this relatively new class of RNA molecules.
Here, we demonstrate that standard expression analysis Affymetrix MOE430 and HG-U133 GeneChips contain hundreds of probe sets that detect NATs. Probe sets carrying a "Negative Strand Matching Probes" annotation in NetAffx were validated using Ensembl by manual and automated approaches. More than 50 % of the 1,113 probe sets with "Negative Strand Matching Probes" on the MOE430 2.0 GeneChip were confirmed as detecting NATs. Expression of selected antisense transcripts as indicated by Affymetrix data was confirmed using strand-specific RT-PCR. Thus, Affymetrix datasets can be mined to reveal information about the regulated expression of a considerable number of NATs. In a correlation analysis of 179 sense-antisense (SAS) probe set pairs using publicly available data from 1637 MOE430 2.0 GeneChips a significant number of SAS transcript pairs were found to be positively correlated.
Standard expression analysis Affymetrix GeneChips can be used to measure many different NATs. The large amount of samples deposited in microarray databases represents a valuable resource for a quantitative analysis of NAT expression and regulation in different cells, tissues and biological conditions.
天然反义转录本(NATs)广泛表达的功能和意义在很大程度上尚不清楚。能够定量评估多个样本中许多不同转录本的NAT表达变化,将有助于我们理解这类相对较新的RNA分子。
在这里,我们证明标准表达分析Affymetrix MOE430和HG-U133基因芯片包含数百个检测NATs的探针组。通过手动和自动方法,使用Ensembl对NetAffx中带有“负链匹配探针”注释的探针组进行了验证。在MOE430 2.0基因芯片上,1113个带有“负链匹配探针”的探针组中,超过50%被确认为可检测NATs。使用链特异性逆转录聚合酶链反应(RT-PCR)证实了Affymetrix数据所显示的选定反义转录本的表达。因此,可以挖掘Affymetrix数据集以揭示大量NATs调控表达的信息。在对179个 sense-antisense(SAS)探针组对进行的相关性分析中,使用来自1637个MOE430 2.0基因芯片的公开数据,发现大量SAS转录本对呈正相关。
标准表达分析Affymetrix基因芯片可用于测量许多不同的NATs。微阵列数据库中存放的大量样本是定量分析不同细胞、组织和生物学条件下NAT表达和调控的宝贵资源。