title: DNAzyme-based probes for telomerase detection in early-stage cancer diagnosis.
abstract: Human telomerase is a polymerase enzyme that adds tandem repeats of DNA (TTAGGG) in the telomeric region to the ends of chromosomes. Since telomerase can be detected in immortalized, but not normal, somatic cells, it has been considered a selective target for cancer chemotherapy. Here, we describe a DNAzyme-based probe to detect the presence of telomerase in cell lysates. Telomerase elongates the primer site on the probe. Subsequent addition of the Pb(II) cofactor activates the DNAzyme, which cleaves the elongated fragment at the RNA site, releasing the probe for repetitive cycling and signal amplification. The cleaved fragment is detected by a reporter molecular beacon. Enzymatic amplification with rapid turnover allows detection of telomerase in the range of 0.1 to 1 μg cell lysate, with a fivefold increase in signal level for cancer cells over normal cells. This probe design can provide a simple, yet rapid and sensitive, measurement of telomerase activity.
authors: Department of Chemistry, Shands Cancer Center and Center for Research at the Bio/Nano Interface, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, FL 32611-7200, USA.
authors: Hui
authors: H
authors: Wang
authors: Michael J
authors: MJ
authors: Donovan
authors: Ling
authors: L
authors: Meng
authors: Zilong
authors: Z
authors: Zhao
authors: Youngmi
authors: Y
authors: Kim
authors: Mao
authors: M
authors: Ye
authors: Weihong
authors: W
authors: Tan
mesh_terms: D004247:DNA / Q000737:chemistry; D021881:DNA, Catalytic / Q000378:metabolism*; D042241:Early Diagnosis; D006801:Humans; D009369:Neoplasms / Q000175:diagnosis*; D012313:RNA / Q000737:chemistry; D012091:Repetitive Sequences, Nucleic Acid; D019098:Telomerase / Q000378:metabolism*
chemical_list: D021881:DNA, Catalytic; D012313:RNA; D004247:DNA; D019098:Telomerase