Functional microRNA screening using a comprehensive lentiviral human microRNA expression library.

Functional microRNA screening using a comprehensive lentiviral human microRNA expression library.
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使用全面的慢病毒人MicroRNA表达式库进行功能性microRNA筛选。

DOI:
10.1186/1471-2164-12-546
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发表时间:
2011-11-03
期刊:
影响因子:
4.4
通讯作者:
Cuppen E
Cuppen E
中科院分区:
生物学2区
文献类型:
--
作者:
Poell JB;van Haastert RJ;Cerisoli F;Bolijn AS;Timmer LM;Diosdado-Calvo B;Meijer GA;van Puijenbroek AA;Berezikov E;Schaapveld RQ;Cuppen E

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MicroRNA (miRNA) 是一类小调节 RNA,其靶向信使 RNA (mRNA) 中的序列以抑制其蛋白质输出。剖析 miRNA 功能的复杂性仍然具有挑战性,因为 miRNA 预计有数千个靶点,而 mRNA 可以被数十个 miRNA 靶向。为了系统地解决 miRNA 的生物学功能,我们构建并验证了一个慢病毒 miRNA 表达库,其中包含 660 个当前注释的和 422 个候选人类 miRNA 前体。 miRNA 从其天然基因组骨架表达,确保生理加工。文库的阵列布局使其成为高通量筛选的理想选择,但也允许集中筛选和命中挑选。我们在短期和长期测定中证明了其功能,并且能够证实先前描述的经过充分研究的 miRNA 的结果。通过 miRNA 表达库,我们为系统阐明 miRNA 功能提供了一个多功能工具。
MicroRNAs (miRNAs) are a class of small regulatory RNAs that target sequences in messenger RNAs (mRNAs) to inhibit their protein output. Dissecting the complexities of miRNA function continues to prove challenging as miRNAs are predicted to have thousands of targets, and mRNAs can be targeted by dozens of miRNAs. To systematically address biological function of miRNAs, we constructed and validated a lentiviral miRNA expression library containing 660 currently annotated and 422 candidate human miRNA precursors. The miRNAs are expressed from their native genomic backbone, ensuring physiological processing. The arrayed layout of the library renders it ideal for high-throughput screens, but also allows pooled screening and hit picking. We demonstrate its functionality in both short- and long-term assays, and are able to corroborate previously described results of well-studied miRNAs. With the miRNA expression library we provide a versatile tool for the systematic elucidation of miRNA function.
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