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利用任意引物PCR进行RNA指纹分析对RNA丰度调节因子之间的相互作用进行表征。

Interactions among regulators of RNA abundance characterized using RNA fingerprinting by arbitrarily primed PCR.

作者信息

McClelland M, Ralph D, Cheng R, Welsh J

机构信息

California Institute of Biological Research, La Jolla 92037.

出版信息

Nucleic Acids Res. 1994 Oct 25;22(21):4419-31. doi: 10.1093/nar/22.21.4419.

Abstract

Using RNA fingerprinting by arbitrarily primed PCR it is possible to infer convergent transcript regulatory pathways from the coordinate behavior of subsets of anonymous transcripts without cloning any genes. The number of transcripts in each response category can be estimated. The same may be true for differential display. We demonstrate these claims by treating a cell line with two known modulators of RNA abundance, transforming growth factor-beta (TGF beta) and cycloheximide (CX), used together and alone. The responses of over 1700 anonymous transcripts were monitored under these three conditions and in an untreated control. Eight of the twenty-seven [3(3)] possible transcript response categories were observed among 86 differentially expressed transcripts. For example, CX stabilizes or induces as many as 2.7% of transcripts of which about one third do not accumulate when TGF beta is also present. This intersection may reflect CX stabilization or induction of an important class of RNAs that otherwise usually have short half-lives. We predict that RNAs in this class constitute the majority of transcripts targeted for rapid down regulation in response to TGF beta and perhaps most other natural transcriptional modulators.

摘要

利用任意引物PCR进行RNA指纹分析,无需克隆任何基因,就有可能从无名转录本子集的协同行为推断出趋同的转录调控途径。可以估计每个反应类别中的转录本数量。差异显示可能也是如此。我们通过用两种已知的RNA丰度调节剂——转化生长因子-β(TGF-β)和环己酰亚胺(CX)单独或联合处理细胞系来证明这些说法。在这三种条件下以及未处理的对照中监测了1700多个无名转录本的反应。在86个差异表达的转录本中观察到了27种[3(3)]可能的转录本反应类别中的8种。例如,CX可稳定或诱导多达2.7%的转录本,当同时存在TGF-β时,其中约三分之一的转录本不会积累。这种交集可能反映了CX对一类重要RNA的稳定或诱导作用,否则这些RNA通常半衰期较短。我们预测,这类RNA构成了响应TGF-β以及可能大多数其他天然转录调节剂而被快速下调的转录本的大部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cc0/308475/5717b558547a/nar00045-0074-a.jpg

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