Polony multiplex analysis of gene expression (PMAGE) in mouse hypertrophic cardiomyopathy
Polony multiplex analysis of gene expression (PMAGE) in mouse hypertrophic cardiomyopathy
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DOI：
10.1126/science.1137325
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发表时间：
2007-06-08
期刊：
影响因子：
56.9
通讯作者：
Seidman, J. G.
中科院分区：
文献类型：
--
作者：
Kim, Jae Bum;Porreca, Gregory J.;Seidman, J. G.
我们描述了一种敏感的mRNA分析技术，PMAGE（用于“基因表达的多态分析”），它检测到每三个细胞中只有一个转录物的信使rna （mRNA）。PMAGE结合了一种改进的基于连接的方法，对来自单个mRNA分子的14个核苷酸标签进行测序。每个mRNA分子的一个序列标签被扩增到一个单独的1微米珠子上，标记为聚合酶集落或poloni，大约500万个poloni被排列在一个流动池中进行平行测序。使用PMAGE，我们在携带致病突变的小鼠中确定了在肥厚性心肌病病理表现之前的早期转录变化。PMAGE提供了心脏mrna的全面概况，包括编码信号分子和可能参与疾病发病机制的转录因子的低丰度mrna。
We describe a sensitive mRNA profiling technology, PMAGE ( for "polony multiplex analysis of gene expression"), which detects messenger RNAs ( mRNAs) as rare as one transcript per three cells. PMAGE incorporates an improved ligation-based method to sequence 14-nucleotide tags derived from individual mRNA molecules. One sequence tag from each mRNA molecule is amplified onto a separate 1-micrometer bead, denoted as a polymerase colony or polony, and about 5 million polonies are arrayed in a flow cell for parallel sequencing. Using PMAGE, we identified early transcriptional changes that preceded pathological manifestations of hypertrophic cardiomyopathy in mice carrying a disease-causing mutation. PMAGE provided a comprehensive profile of cardiac mRNAs, including low-abundance mRNAs encoding signaling molecules and transcription factors that are likely to participate in disease pathogenesis.