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

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.

SUBMITTER: Alkailani MI

PROVIDER: S-EPMC11320832 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature

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

Alkailani Maisa I MI

Biology methods & protocols 20240528 1

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 des ...[more]

PMID: 39139996

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