Under this grant we propose to develop and test a new class of fluorescent oligonucleotide probes based on a novel high performance lanthanide chelate design. By virtue of their long fluorescence emission lifetimes, lanthanide reporter oligonucleotide process will offer fundamentally improved performance over conventional fluorescent probes. The lanthanide chelate complex we will evaluate as a DNA reporter, is a new design based on a quarter of isopthalamide ligands. Lanthanide complexes prepared from this chelate are stable while they capture and emit light more efficiently than previous designs, potentially allowing assays with greater S/N, and thus greater sensitivity. Our primary objective under phase one will be to synthesize oligonucleotide probes based on lanthanide complex and confirm the expected performance characteristics of the probes within a variety of hybridization assays. Extension of the technology will include development of methodology allowing automated solid phase synthesis of lanthanide complex-oligonucleotide conjugates, and the application of labeled probes to gene quantitation and DNA array mediated assays.
Luminescent lanthanide complexes have great potential as oligonucleotide probe reporter groups. Under this grant, advanced lanthanide complexes will be developed and adapted to oligonucleotide synthesis methodology. As a result the practical and economic synthesis of lanthanide oligo probes will become possible, and their application to genomic assays can begin.
| Johansson, Mary Katherine; Cook, Ronald M; Xu, Jide et al. (2004) Time gating improves sensitivity in energy transfer assays with terbium chelate/dark quencher oligonucleotide probes. J Am Chem Soc 126:16451-5 |
