Northern blotting of endogenous full-length human-specific LINE-1 RNA.

IF 2.5 Q3 BIOCHEMICAL RESEARCH METHODS Biology Methods and Protocols Pub Date : 2024-05-28 eCollection Date: 2024-01-01 DOI:10.1093/biomethods/bpae036
Maisa I Alkailani
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Abstract

LINE-1 belongs to a family of DNA elements that move to new locations in the genome in a process called "retrotransposition." This is achieved by a copy-and-paste mechanism with the aid of an RNA intermediate. The full-length LINE-1 is responsible for most retrotransposition activity in the human genome. Detecting the active LINE-1 RNA at the endogenous level is challenging due to its small percentage among inactive copies and its different forms of transcripts. Here, we describe a method of designing RNA probes to detect active LINE-1 by northern blotting and use optimized conditions and tools to make the detection practical. This method uses a classical long RNA probe and provides an alternative way to detect LINE-1 RNA using multiple short RNA probes.

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内源性全长人类特异性 LINE-1 RNA 的 Northern 印迹。
LINE-1属于DNA元件家族的一员,它们通过一种叫做 "逆转录 "的过程转移到基因组中的新位置。这是通过一种复制粘贴机制,借助 RNA 中间体实现的。全长 LINE-1 负责人类基因组中的大部分逆转录活动。由于 LINE-1 RNA 在非活性拷贝中所占比例很小,而且其转录本形式各异,因此在内源性水平检测活性 LINE-1 RNA 是一项挑战。在这里,我们介绍了一种设计 RNA 探针的方法,通过北印迹法检测活性 LINE-1,并使用优化的条件和工具使检测切实可行。该方法使用经典的长 RNA 探针,并提供了一种使用多个短 RNA 探针检测 LINE-1 RNA 的替代方法。
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来源期刊
Biology Methods and Protocols
Biology Methods and Protocols Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
3.80
自引率
2.80%
发文量
28
审稿时长
19 weeks
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