A Semisynthetic Bioluminescence Sensor for Ratiometric Imaging of Metal Ions In Vivo Using DNAzymes Conjugated to An Engineered Nano-Luciferase |
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Authors: | Dr. Mengyi Xiong Dr. Yuting Wu Dr. Gezhi Kong Whitney Lewis Dr. Zhenglin Yang Hanxiao Zhang Li Xu Ying Liu Qin Liu Prof. Xuhua Zhao Prof. Xiao-Bing Zhang Prof. Yi Lu |
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Affiliation: | 1. Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, 410082 Changsha, Hunan, P. R. China;2. Department of Chemistry, University of Texas at Austin, 78712 Austin, TX, USA;3. Department of Biochemistry and Molecular Biology, Shanxi Medical University, 030001 Taiyuan, Shanxi, P. R. China |
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Abstract: | DNA-based probes have gained significant attention as versatile tools for biochemical analysis, benefiting from their programmability and biocompatibility. However, most existing DNA-based probes rely on fluorescence as the signal output, which can be problematic due to issues like autofluorescence and scattering when applied in complex biological materials such as living cells or tissues. Herein, we report the development of bioluminescent nucleic acid (bioLUNA) sensors that offer laser excitation-independent and ratiometric imaging of the target in vivo. The system is based on computational modelling and mutagenesis investigations of a genetic fusion between circular permutated Nano-luciferase (NLuc) and HaloTag, enabling the conjugation of the protein with a DNAzyme. In the presence of Zn2+, the DNAzyme sensor releases the fluorophore-labelled strand, leading to a reduction in bioluminescent resonance energy transfer (BRET) between the luciferase and fluorophore. Consequently, this process induces ratiometric changes in the bioluminescent signal. We demonstrated that this bioLUNA sensor enabled imaging of both exogenous Zn2+ in vivo and endogenous Zn2+ efflux in normal epithelial prostate and prostate tumors. This work expands the DNAzyme sensors to using bioluminescence and thus has enriched the toolbox of nucleic acid sensors for a broad range of biomedical applications. |
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Keywords: | Bioluminescence DNAzyme Nucleic Acid Sensor In Vivo Imaging Protein Engineering |
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