RNA-based cooperative protein labeling that permits direct monitoring of the intracellular concentration change of an endogenous protein

Abstract Imaging the dynamics of proteins in living cells is a powerful means for understanding cellular functions at a deeper level. Here, we report a versatile method for spatiotemporal imaging of specific endogenous proteins in living mammalian cells. The method employs a bifunctional aptamer cap...

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Bibliographic Details
Published in:	Nucleic acids research Vol. 49; no. 22; p. e132
Main Authors:	Pe, Kathleen Beverly Alog, Yatsuzuka, Kenji, Hakariya, Hayase, Kida, Tomoki, Katsuda, Yousuke, Fukuda, Masatora, Sato, Shin-ichi
Format:	Journal Article
Language:	English
Published:	England Oxford University Press 16-12-2021
Subjects:	Actins - analysis Aptamers, Nucleotide - chemistry Fluorescent Dyes HeLa Cells Humans Methods Online Molecular Imaging - methods Optical Imaging - methods Proteins - analysis RNA - chemistry Time-Lapse Imaging
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Summary:	Abstract Imaging the dynamics of proteins in living cells is a powerful means for understanding cellular functions at a deeper level. Here, we report a versatile method for spatiotemporal imaging of specific endogenous proteins in living mammalian cells. The method employs a bifunctional aptamer capable of selective protein recognition and fluorescent probe-binding, which is induced only when the aptamer specifically binds to its target protein. An aptamer for β-actin protein preferentially recognizes its monomer forms over filamentous forms, resulting in selective G-actin staining in both fixed and living cells. Through actin-drug treatment, the method permitted direct monitoring of the intracellular concentration change of endogenous G-actin. This protein-labeling method, which is highly selective and non-covalent, provides rich insights into the study of spatiotemporal protein dynamics in living cells. Graphical Abstract Graphical Abstract A convenient and highly-selective protein-labeling technology permitted direct monitoring of intracellular concentration change of endogenous G-actin proteins, which is normally very hard to distinguish from F-actin by live-cell staining.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0305-1048 1362-4962
DOI:	10.1093/nar/gkab839