Method for measuring in vivo inhibition of intracellular RNase
US10663456B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 6, 2016 |
| Grant date | May 26, 2020 |
| Priority date | — |
| Expiry date | Jul 6, 2036 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/582
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present invention provides means for evaluating harmfulness of a chemical substance before occurrence of cell death, that is, more quickly and sensitively compared to conventional methods wherein the remaining viable cell count is determined using a reductive coloring reagent after a period of time required for occurrence of cell death due to the chemical substance. A double-stranded RNA probe comprising an RNA strand labeled with a fluorescent dye A that emits fluorescence, and an RNA strand labeled with a fluorescent dye B that quenches emission from a fluorescent dye in the vicinity thereof, wherein the fluorescence is quenched in a double-stranded state due to occurrence of fluorescence resonance energy transfer (FRET) between the two kinds of fluorescent dyes. When the probe is introduced into a cell and the cell is in a normal state, the double-stranded RNA is quickly degraded by activity of intracellular ribonuclease to cause cancellation of the FRET state, allowing fluorescence emission of the fluorescent dye A. On the other hand, in cases where the cell is harmfully influenced, activity of intracellular ribonuclease is suppressed, so that the double-stranded RNA remain…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.