DNA detection and signal amplification via an engineered allosteric enzyme.
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.
Saghatelian
Alan
A
Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, USA.
Guckian
Kevin M
KM
Thayer
Desiree A
DA
Ghadiri
M Reza
MR
D001409:Bacillus cereus / Q000201:enzymology* / Q000235:genetics; D004247:DNA / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000737:chemistry; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry*; D012815:Signal Processing, Computer-Assisted
D004277:DNA, Single-Stranded; D009842:Oligopeptides; D011480:Protease Inhibitors; D004247:DNA; D008666:Metalloendopeptidases