Identification of novel long noncoding RNA transcripts in male germ cells.
Identification of novel long noncoding RNA transcripts in male germ cells.
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DOI：
10.1007/978-1-61779-436-0_9
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发表时间：
2012
期刊：
影响因子：
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通讯作者：
Rennert, Owen M
中科院分区：
文献类型：
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作者：
Lee, Tin-Lap;Xiao, Amy;Rennert, Owen M
来自这些研究的新证据表明，男性生殖细胞转录组比以前设想的更复杂。除了蛋白质编码基因外，转录组还编码大量的非蛋白质编码转录物。这些非编码（nc） rna似乎参与各种细胞活动，从简单的家务管理到复杂的调节功能。一类被称为长链ncRNAs （lncRNAs）的ncR-NAs最近被证明在脑和胚胎干细胞发育过程中以发育调节的方式表达。该方案旨在利用基因表达序列分析（SAGE）数据预测和鉴定潜在的lncRNA候选物。我们还说明了如何通过实时PCR和Northern Blot的表达分析来验证潜在的lncrna。使用我们先前建立的男性生殖细胞SAGE数据库（GermSAGE）鉴定男性生殖细胞中潜在的lncRNA候选者。
Emerging evidence from these studies suggested that the male germ cell transcriptome is more complex than previously envisioned. In addition to protein-coding genes, the transcriptome also encodes a significant number of nonprotein-coding transcripts. These noncoding (nc) RNAs appear to be involved in a variety of cellular activities, ranging from simple housekeeping to complex regulatory functions. A class of ncR-NAs known as long ncRNAs (lncRNAs) were recently shown to be expressed in a developmentally regulated manner during brain and embryonic stem cell development. This protocol aims to predict and identify potential lncRNA candidates using Serial Analysis of Gene Expression (SAGE) data. We also illustrate how to validate the potential lncRNAs by expression analyses using real-time PCR and Northern Blot. Potential lncRNA candidates in male germ cells are identified using our previously established male germ cell SAGE database (GermSAGE).