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A Supercharged Fluorescent Protein as a Versatile Probe for Homogeneous DNA Detection and Methylation Analysis

Authors

  • Chunyang Lei,

    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
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  • Dr. Yan Huang,

    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
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  • Prof. Dr. Zhou Nie,

    Corresponding author
    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
    • State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)===

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  • Jun Hu,

    1. Pathology Department of Hunan Provincial Tumor Hospital/ the Affiliated Tumor Hospital of Xiangya Medical School, Central South University, Changsha, Hunan 410013 (China)
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  • Lijun Li,

    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
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  • Guoyan Lu,

    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
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  • Yitao Han,

    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
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  • Prof. Dr. Shouzhuo Yao

    1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China)
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  • We are grateful to the National Natural Science Foundation of China (Nos. 21222507, 21175036, 21235002, and 21190044), the National Basic Research Program of China (973 Program, No. 2011CB911002), and the Foundation for Innovative Research Groups of NSFC (Grant 21221003).

Abstract

Supercharged proteins are a new class of functional proteins with exceptional stability and potent ability to deliver bio-macromolecules into cells. As a proof-of-principle, a novel application of supercharged proteins as a versatile biosensing platform for nucleic acid detection and epigenetics analysis is presented. Taking supercharged green fluorescent protein (ScGFP) as the signal reporter, a simple turn-on homogenous method for DNA detection has been developed based on the polyionic nanoscale complex of ScGFP/DNA and toehold strand displacement. This assay shows high sensitivity and potent ability to detect single-base mismatch. Furthermore, combined with bisulfite conversion, this ScGFP-based assay was further applied to analyze site-specific DNA methylation status of genomic DNA extracted from real human colon carcinoma tissue sample with ultrahigh sensitivity (4 amol methylated DNA).

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