Alkailani Maisa I
College of Health and Life Sciences, Hamad Bin Khalifa University, Qatar Foundation, Doha, P.O. Box 34110, Qatar.
Biol Methods Protoc. 2024 May 28;9(1):bpae036. doi: 10.1093/biomethods/bpae036. eCollection 2024.
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.
LINE-1属于一类DNA元件家族，它们通过一种称为“逆转座”的过程移动到基因组中的新位置。这是通过一种在RNA中间体帮助下的复制粘贴机制实现的。全长LINE-1负责人类基因组中的大多数逆转座活性。由于其在非活性拷贝中所占比例小以及转录本形式不同，在内源水平检测活性LINE-1 RNA具有挑战性。在此，我们描述了一种设计RNA探针通过Northern印迹检测活性LINE-1的方法，并使用优化的条件和工具使检测具有实用性。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。
