A Supercharged Fluorescent Protein as a Versatile Probe for Homogeneous DNA Detection and Methylation Analysis |
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| Authors: | Chunyang Lei Dr. Yan Huang Prof. Dr. Zhou Nie Jun Hu Lijun Li Guoyan Lu Yitao Han Prof. Dr. Shouzhuo Yao |
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| Affiliation: | 1. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, ChangSha 410082 (P.R. China);2. 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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| 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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| Keywords: | bioanalysis DNA DNA methylation supercharged protein toehold strand displacement |
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