Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 53, Issue 32, Pages 8358-8362Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201403615
Keywords
bioanalysis; DNA; DNA methylation; supercharged protein; toehold strand displacement
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Funding
- National Natural Science Foundation of China [21222507, 21175036, 21235002, 21190044]
- National Basic Research Program of China (973 Program) [2011CB911002]
- Foundation for Innovative Research Groups of NSFC [21221003]
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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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