An approach toward SNP detection by modulating the fluorescence of DNA-templated silver nanoclusters.
A rapid, homogeneous, and in-situ labelable format designed to detect single nucleotide polymorphism (SNP) is presented. Fluorescent silver nanoclusters produced following hydride-mediated reduction of Ag(+) bound to a partial double-stranded oligodeoxynucleotide were employed as probes for SNPs. The sensing mechanism is based on the fluorescence enhancement of silver nanoclusters through an irreversible cluster transfer upon hybridization. The key element of our design is modulating the "turn on" mechanism by introducing a competitor that can be displaced in response to a target DNA. In a controlled model system, the fluorescence intensity of silver nanoclusters is approximately 3-fold enhanced upon hybridization with a perfectly matched target DNA, and single-base mismatch detection is achieved within 5min, regardless of the position of mismatches. Human aldehyde dehydrogenase 2 (ALDH2), which is responsible for the oxidation of aldehydes to carboxylic acids, is then utilized as a target for SNP. Upon addition of a perfectly matched sequence, dramatically enhanced fluorescence intensity is observed (ca. 48-fold), and prompt SNP genotyping is accomplished.
Department of Chemistry, Pohang University of Science and Technology (POSTECH), Pohang 790-784, Republic of Korea.
Juhee
J
Park
Jihyun
J
Lee
Changill
C
Ban
Won Jong
WJ
Kim
D000327:Adsorption; D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation*; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D053768:Metal Nanoparticles / Q000737:chemistry*; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012834:Silver / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*
D012834:Silver; D004247:DNA