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Analytical and Bioanalytical Chemistry

, Volume 391, Issue 3, pp 983–991 | Cite as

Investigation of the hybrid molecular probe for intracellular studies

  • Karen Martinez
  • Colin D. Medley
  • Chaoyong James Yang
  • Weihong Tan
Original Paper

Abstract

Monitoring gene expression in vivo is essential to the advancement of biological studies, medical diagnostics, and drug discovery. Adding to major efforts in developing molecular probes for mRNA monitoring, we have recently developed an alternative tool, the hybrid molecular probe (HMP). To optimize the probe, a series of experiments were performed to study the properties of HMP hybridization kinetics and stability. The results demonstrated the potential of the HMP as a prospective tool for use in both hybridization studies and in vitro and in vivo analyses. The HMP has shown no tendency to produce false positive signals, which is a major concern for living cell studies. Moreover, HMP has shown the ability to detect the mRNA expression of different genes inside single cells from both basal and stimulated genes. As an effective alternative to conventional molecular probes, the proven sensitivity, simplicity, and stability of HMPs show promise for their use in monitoring mRNA expression in living cells.

Figure

Hybrid molecular probe (HMP). HMPs consist of two single strands of DNA (green) and a polyethylene glycol (PEG, purple) linker that is used to tether these two sequences together. When a target (orange strand) containing the complementary sequences to both probes at adjacent positions is added, each strand binds to its corresponding target sequence, thus bringing the two fluorophores into close proximity, which allows energy transfer to occur

Keywords

DNA detection Fluorescence probes Molecular beacons Fluorescence resonance energy transfer (FRET) mRNA monitoring 

Notes

Acknowledgment

This work was supported by NIH grants 5R01GM066137 and 5R21CA122648, and a grant from the Center of Excellence in Bio/nano sensors, State of Florida.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Karen Martinez
    • 1
  • Colin D. Medley
    • 1
  • Chaoyong James Yang
    • 1
  • Weihong Tan
    • 1
  1. 1.Center for Research at the Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, UF Genetics Institute and McKnight Brain InstituteUniversity of FloridaGainesvilleUSA

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