A combined immunoprecipitation, mass spectrometric and nucleic acid sequencing approach to determine microRNA-mediated post-transcriptional gene regulatory networks.
A combined immunoprecipitation, mass spectrometric and nucleic acid sequencing approach to determine microRNA-mediated post-transcriptional gene regulatory networks.
复制标题
结合免疫沉淀、质谱和核酸测序方法来确定 microRNA 介导的转录后基因调控网络。
DOI：
10.1093/bfgp/elp050
复制
发表时间：
2010
影响因子：
4
通讯作者：
Tanese,Naoko
中科院分区：
文献类型：
--
作者：
Savas,JeffreyN;Tanese,Naoko
虽然转录起始在基因调控领域引起了最多的关注，但已经清楚的是，包括转录后事件在内的基因表达级联中的其他阶段也处于同样精确的控制之下。短RNA（microRNA，small interfering RNA，Piwi-interacting RNA）在抑制基因表达中发挥重要作用的开创性发现，激发了人们对转录后基因沉默机制的新兴趣。亲和标签和高分辨率串联质谱（MS/MS）的发展大大简化了调节基因表达的蛋白质的分析。此外，DNA微阵列和“第二代”核酸测序（“深度测序”）技术的使用促进了其调控靶标的鉴定。这些技术进步标志着全面了解基因调控网络的重要一步。这篇综述的目的是强调最近的几篇报道，这些报道说明了亲和纯化（免疫沉淀）的价值，然后通过深度测序（AP-MS/Seq）进行质谱蛋白质分析和核酸分析，以检查转录后的mRNA。识别转录后基因调控机器的直接核酸靶标的能力是理解转录后途径对基因表达的贡献的关键的第一步。
While initiation of transcription has attracted the most attention in the field of gene regulation, it has become clear that additional stages in the gene expression cascade including post-transcriptional events are under equally exquisite control. The seminal discovery that short RNAs (microRNA, small interfering RNA, Piwi-interacting RNA), play important roles in repressing gene expression has spurred a rush of new interest in post-transcriptional gene silencing mechanisms. The development of affinity tags and high-resolution tandem mass spectrometry (MS/MS) has greatly simplified the analysis of proteins that regulate gene expression. Further, the use of DNA microarrays and ‘second generation’ nucleic acid sequencing (‘deep sequencing’) technologies has facilitated the identification of their regulatory targets. These technological advancements mark a significant step towards a comprehensive understanding of gene regulatory networks. The purpose of this review is to highlight several recent reports that illustrate the value of affinity-purification (immunoprecipitation) followed by mass spectrometric protein analysis and nucleic acid analysis by deep sequencing (AP-MS/Seq) to examine mRNA after it has been transcribed. The ability to identify the direct nucleic acid targets of post-transcriptional gene regulatory machines is a critical first step towards understanding the contribution of post-transcriptional pathways on gene expression.