Method of measuring oligonucleotide decomposing activity
AssigneeHamamatsu Photonics K.K.
InventorsHisatoshi Uchiyama, Masaki Jibu, Kenichi Hirano, Kazunari Taira
Life of this patent
Estimating the term…
Abstract
The present invention provides an in situ monitoring of the decomposing activity against oligonucleotides target in a biological tissue. A single-chain oligonucleotide target comprises an appropriate number of nucleic acid bases to be examined with an energy donor and an energy acceptor respectively at its 5'- and 3'-terminals. Monitoring of the fluorescence changes of the target after injection into a biological tissue, particularly the fluorescent resonance energy transfer(FRET) phenomena between the energy donor and acceptor, indicates whether the oligonucleotide is not decomposed yet.
Claims10
A method of measuring decomposition of a single-stranded oligonucleotide by endonuclease activity, wherein the oligonucleotide comprises an energy donor group and an energy acceptor group positioned sufficiently close so as to have overlapping energy spectrums, and a restriction site recognizable by an endonuclease, which restriction site is positioned between the energy donor group and the energy acceptor group,
- which method comprises the steps of
- (i) detecting a resonance energy transfer between the energy donor group and the energy acceptor group;
- and
- (ii) detecting a loss in the resonance energy transfer, which loss indicates decomposition of the oligonucleotide by endonuclease activity at the restriction site.
Dependencies are inferred from the claim text.