Selection and Application of Light-up Nucleic Acid Aptamers

doi:10.13560/j.cnki.biotech.bull.1985.2021-1183

Abstract

Abstract:

Light-up nucleic acid aptamers（LNAs）are a class of functional nucleic acids that can specifically bind to target molecules and enhance the luminescence performance of target molecules. LNAs have the advantages of simple composition,stable structure,and short cell expression time,thus they show great potential in the field of cell imaging,substance detection and other sensing fields. With the in-depth research on the in-situ analysis of intracellular genes,the deficiencies of the existing intracellular imaging technology have gradually appeared,and the LNAs imaging system has emerged. This review sorts out and summarizes the selecting methods of different types of LNAs,analyzes the frontier applications of LNAs from both intracellular and in vitro aspects,points out the deficiencies of the current LNAs research and the sensing system,and looks forward to the huge development prospects of LNAs in cell-free sensing,aiming to promote the LNAs system becoming a powerful tool for the research of intracellular and extracellular sensing.

Key words: light-up nucleic acid aptamers, fluorophore, RNA imaging, functional nucleic acids

ZHOU Zi-qi, ZHANG Yang-zi, LAN Xin-yue, LIU Yang-er, ZHU Long-jiao, XU Wen-tao. Selection and Application of Light-up Nucleic Acid Aptamers[J]. Biotechnology Bulletin, 2022, 38(5): 240-247.

Figures/Tables 3

Fig. 1 In vitro selection of LNAs using SELEX and other techniques

Fig. 2 Application of LNAs system in living cells A: Monitoring the formation of (CGG)60-Spinach 2 foci in transiently transfected COS-7 cell. B: Application of m6A-modified Broccoli as a fluorometric substrate in the FTO assay. C: Imaging SAM in living cells with RNA aptamer and Spinach. D: Assembly and sensing of aptamer in split Spinach

Fig. 3 Application of LNAs system in vitro A: Schematic diagram of fluorescent bivalent metal ion sensor based on riboswitch, secondary structure of czcD-2 and anaerobic titration experiment. B: Schematic representation of the in-gel detection of vegetable aptamer-tagged RNAs and their cleavage products. C: Sensing equipment and principle of ROSALIND system

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