IVT-SAPAS: Low-Input and Rapid Method for Sequencing Alternative Polyadenylation Sites.
IVT-SAPAS: Low-Input and Rapid Method for Sequencing Alternative Polyadenylation Sites.
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IVT-SAPAS：用于对替代多聚腺苷酸化位点进行测序的低输入快速方法。
DOI：
10.1371/journal.pone.0145477
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Xu A
中科院分区：
文献类型：
--
作者：
Fu Y;Ge Y;Sun Y;Liang J;Wan L;Wu X;Xu A
基因转录与选择性多聚腺苷酸化(APA)位点结合导致了不同的mRNA异构体，其可能编码不同的蛋白质或含有不同的3‘UTRs。APA在发育、免疫反应和癌症等多种生理过程中对基因表达网络起着重要的调控作用。为了应用高通量测序，已经开发了几种用于APA研究的文库构建方法。然而，现有方法对高投入RNA的要求和耗时的性质限制了对难以获得的样品的APA研究。在这里，我们描述了一种基于我们的SAPAS的结合体外转录(IVT)和磁珠纯化的新方法。新的IVT-SAPAS为低输入样本的APA文库构建提供了一种快速、高度并行的方法，为APA的研究提供了一种新的稳健的方法。
Gene transcribing with alternative polyadenylation (APA) sites leads to mRNA isoforms, which may encode different proteins or harbor different 3'UTRs. APA plays an important role in regulating gene expression network among various physiological processes, such as development, immune responses and cancer. Several methods of library construction for APA study have been developed to apply high-throughput sequencing. However, the requirement of high-input RNA and time-consuming nature of the current methods limited the studies of APA for the samples difficult to obtain. Here, we describe a new method based on our SAPAS in combining in vitro transcription (IVT) and magnetic beads purification. The new IVT-SAPAS provides a rapid and high-parallel procedure for APA library construction with low-input sample, which may be a new robust approach for studying APA.