Novel labeling method for messenger RNA and circular RNA based on bimolecular fluorescence complementation

Abstract

RNA molecules are an important member of an intracellular genetic information delivery system. Conventional RNA labeling methods cannot simultaneously have the characteristics of living cells, no invasion, high temporal and spatial resolution, single molecules, etc., so a novel method for labeling and tracking RNA need to be developed. The invention discloses a novel labeling method for messengerRNA and circular RNA based on self-ligation label bimolecular fluorescence complementation. A RNA aptamer subsystem is cooperatively used, aptamers are inserted at proper positions in the RNA, and twoparts of a self-ligation label are respectively fused with the binding proteins of the RNA aptamers. When the fusion proteins do not bind to RNA, RNA molecules do not fluoresce; when and only when the fusion proteins bind to RNA, the RNA molecules fluoresce, so the fluorescence background of an intracellular RNA marker is reduced. The novel RNA labeling method based on self-ligation label bimolecular fluorescence complementation can be used for long-term interference-free single-molecule imaging observation and tracking of messenger RNA and circular RNA of living cells and fixed cells, and has good application prospects in cell biology.

Description

technical field [0001] The present invention relates to a novel intracellular messenger RNA and circular RNA labeling method and its application in cell biology. in the field of cell biology. Background technique [0002] Ribonucleic acid (RNA) molecules are carriers of genetic information present in biological cells and some viruses and viroids. A ribonucleotide monomer molecule is composed of a molecule of phosphate, ribose sugar and base. There are four main bases of ribonucleotide monomers, namely adenine (A), guanine (G), cytosine (C), and uracil (U). RNA is a long-chain polymer formed by the condensation of ribonucleotides through phosphodiester bonds. The central dogma states that DNA is transcribed to form RNA, and RNA is translated into protein to function. RNA is a single strand formed by transcription based on a strand of DNA as a template and based on the principle of complementary base pairing. Its main function is to realize the transfer of genetic informat...

AI Technical Summary

Problems solved by technology

Since each circular RNA and its corresponding linear isoform are spliced ​​from the same precursor RNA, circular RNA, linear RNA and precursor RNA share part of the nucleotide sequence, and it is difficult to target the sequence specific markers to distinguish the three

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