Gene Manipulation with Micro RNAs at Single-Human Cancer Cell.
Gene Manipulation with Micro RNAs at Single-Human Cancer Cell.
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在单个人类癌细胞中使用 Micro RNA 进行基因操作。
DOI：
10.1007/978-1-4939-7601-0_18
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Zhong,JiangF
中科院分区：
文献类型：
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作者：
Stucky,Andres;Chen,Xuelian;Zhong,JiangF
微小RNA（miRNA）是由较长的前体RNA转录物加工而成的小RNA，其可以折叠回自身以形成沃森-克里克配对发夹结构。一旦从较长的分子中加工出来，小RNA就太短了，无法编码蛋白质，但可以发挥其他非常重要的作用，如基因调控。RNA干扰现象最初是由Napoli和Jorgensen在转基因矮牵牛花中观察到的，其中在引入属于类黄酮生物合成途径的查耳酮合酶（CHS）的转基因后观察到基因抑制。miRNAs最初被发现是因为它们在发育中的作用，但很快就发现它们在癌症中也有因果作用。miRNA也可用于操纵基因以研究致癌作用。我们实验室的单细胞转录组分析研究表明，致癌作用往往是一个分子途径的多个成员功能障碍的结果。在这里，我们描述了一个协议，以操纵多个癌症相关基因在一个单一的人类细胞，以调查如何多个基因在致癌过程中相互作用。
Micro RNAs (miRNAs) are small RNAs processed from longer precursor RNA transcripts that can fold back on themselves to form Watson-Crick paired hairpin structures. Once processed from the longer molecule, the small RNA is much too short to code for proteins but can play other very important roles, like gene regulation. The phenomenon of RNA interference was initially observed by Napoli and Jorgensen in transgenic petunia flowers, where gene suppression was observed after introducing a transgene of chalcone synthase (CHS) belonging to the flavonoid biosynthesis pathway. miRNAs were first discovered for their roles in development but it has quickly become evident that they have causal roles in cancer as well. miRNA can also be used to manipulate genes for the investigation of carcinogenesis. Single-cell transcriptome profiling studies in our laboratory suggest that carcinogenesis often is the result of the malfunction of multiple members of a molecular pathway. Here, we describe a protocol to manipulate multiple cancer-related genes in a single human cell to investigate how multiple genes interact during carcinogenesis.