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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 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.

SUBMITTER: Pe KBA 

PROVIDER: S-EPMC8682759 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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

Pe Kathleen Beverly Alog KBA   Yatsuzuka Kenji K   Hakariya Hayase H   Kida Tomoki T   Katsuda Yousuke Y   Fukuda Masatora M   Sato Shin-Ichi SI  

Nucleic acids research 20211201 22


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 it  ...[more]

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