Pyrophosphate anion (PPi) is involved in many biological processes that include ATP hydrolysis, DNA and RNA polymerizations, and enzymatic reactions. The anion is now emerging as a specific signaling molecule in bones and cells. The level of PPi in human tissue is directly related to various enzymatic activities, whose abnormal activities can cause diseases such as heart disease and Alzheimer disease. The detection of pyrophosphate is also an essential component in the commercial DNA pyrosequencing, which provides the necessary genetic profile for hospital patients. Although the genome-based medicine can dramatically increase the efficiencies in the patient treatment, the current high cost associated with the DNA sequencing prevents its routine applications. The development of low cost method for PPi detection, therefore, has a profound impact on the disease studies and DNA sequencing. The current method to detect PPi is based on a complicated enzymatic process, involving two enzymatic reactions occurring in sequence. A simple fluorescent detection of PPi could greatly reduce the cost for this step, which ultimately leads to price reduction for DNA sequencing. The proposed research is dedicated to find a simple solution for non-enzymatic detection of PPi. Our approach is to synthesize a chemical sensor that integrates a binuclear Zn(II)-Zn(II) core into 2-(2'- hydroxyphenyl)benzoxazole structure. The PPi binding to the sensor will trigger the excited state intramolecular proton transfer (ESIPT), thereby leading to a large optical response. Preliminary results show that the new pyrophosphate sensors can provide a fluorescence-based bioassay that greatly simplifies the current pyrophosphate detection scheme. The developed sensor has the potential to be used in DNA pyrosequencing.
Detection of pyrophosphate anion is an important element in DNA pyrosequencing, which currently uses the complicated enzymatic reactions and provides essential information for genome-based medicine. The proposed research focuses on the development of highly selective pyrophosphate sensor, which can greatly simplify the costly enzymatic detection scheme for DNA pyrosequencing and therefore reduce the DNA sequencing cost. Success in the proposed direction will have an important impact in achieving economically acceptable human genome analysis in a clinical setting.
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