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DNA Detection and Signal Amplification via an Engineered Allosteric Enzyme

2002; American Chemical Society; Volume: 125; Issue: 2 Linguagem: Inglês

10.1021/ja027885u

1943-2984

Alan Saghatelian, Kevin M. Guckian, Desiree A. Thayer, M. Reza Ghadiri,

CRISPR and Genetic Engineering

Rapid, sensitive, and sequence-specific DNA detection can be achieved in one step using an engineered intrasterically regulated enzyme. The semi-synthetic inhibitor-DNA-enzyme (IDE) construct (left) rests in the inactive state but upon exposure to a complementary DNA sequence undergoes a DNA hybridization-triggered allosteric enzyme activation (right). The ensuing rapid substrate turnover provides the built-in signal amplification mechanism for detecting approximately 10 fmol DNA in less than 3 min under physiological conditions.