Knockout Cell lines | study of non-coding RNA

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Published on: August 13, 2020

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With the progress of studying microRNA, it is revealed that ncRNA plays an important role in post-transcriptional regulation, cell growth, differentiation and proliferation. MicroRNA, circRNA and lncRNA are the most commonly studied ncRNA. Since 2018, over 60,000 publications are related to non-coding RNA.

What are the methods to study the function of non-coding RNA? And how to choose the best method for your research?

Overexpression, RNA mimics, and RNA agonists can be used for functional acquisition verification. For the study of functional deficiency, RNAi, RNA inhibitors, antagonists, antisense oligonucleotides (ASO), promoter knockout, locus knockout are commonly used.

RNAi cannot silence non-coding RNA like some circRNA and lncRNA. RNAi plays a role in the post-transcriptional level, that is to say, RNA must be located in the cytoplasm to be interfered. But lncRNA and circRNA is selectively distributed in the nucleus and cytoplasm.
RNA inhibitors and antagonists are mainly used in the research of microRNA, but not in the research of circRNA and lncRNA.
ASO cannot completely prevent RNA transcription and interfere with expression at the DNA level. In addition, using ASO method cannot generate stable transgenic cell lines.

Locus knockout and promoter knockout are the most effective methods to study the function of long non-coding RNA.

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