Inert Pepper aptamer-mediated endogenous mRNA recognition and imaging in living cells

Author:

Wang Qi1ORCID,Xiao Feng1,Su Haomiao12ORCID,Liu Hui1,Xu Jinglei1,Tang Heng1,Qin Shanshan1,Fang Zhentian1,Lu Ziang1,Wu Jian3,Weng Xiaocheng1ORCID,Zhou Xiang14ORCID

Affiliation:

1. College of Chemistry and Molecular Sciences, Key Laboratory of Biomedical Polymers-Ministry of Education, Wuhan University , Luojiashan Street, Wuchang District, Wuhan, HuBei 430072, PR China

2. Department of Chemistry, Yale University , 225 Prospect Street , New Haven , CT 06520, USA

3. School of Medicine, Wuhan University , Luojiashan Street, Wuchang District , Wuhan , HuBei 430072, PR China

4. The Institute of Advanced Studies, Wuhan University , Luojiashan Street, Wuchang District , Wuhan , HuBei 430072, PR China

Abstract

Abstract The development of RNA aptamers/fluorophores system is highly desirable for understanding the dynamic molecular biology of RNAs in vivo. Peppers-based imaging systems have been reported and applied for mRNA imaging in living cells. However, the need to insert corresponding RNA aptamer sequences into target RNAs and relatively low fluorescence signal limit its application in endogenous mRNA imaging. Herein, we remolded the original Pepper aptamer and developed a tandem array of inert Pepper (iPepper) fluorescence turn-on system. iPepper allows for efficient and selective imaging of diverse endogenous mRNA species in live cells with minimal agitation of the target mRNAs. We believe iPepper would significantly expand the applications of the aptamer/fluorophore system in endogenous mRNA imaging, and it has the potential to become a powerful tool for real-time studies in living cells and biological processing.

Funder

National Natural Science Foundation of China

Publisher

Oxford University Press (OUP)

Subject

Genetics

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