Invention Grant
- Patent Title: Method for measuring in vivo inhibition of intracellular RNase
-
Application No.: US15746570Application Date: 2016-07-06
-
Publication No.: US10663456B2Publication Date: 2020-05-26
- Inventor: Hidenori Tani , Masaki Torimura , Hiroaki Sato
- Applicant: NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
- Applicant Address: JP Tokyo
- Assignee: NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
- Current Assignee: NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
- Current Assignee Address: JP Tokyo
- Agency: Wenderoth, Lind & Ponack, L.L.P.
- Priority: com.zzzhc.datahub.patent.etl.us.BibliographicData$PriorityClaim@36baca58
- International Application: PCT/JP2016/070036 WO 20160706
- International Announcement: WO2017/018145 WO 20170202
- Main IPC: C07H21/04
- IPC: C07H21/04 ; G01N33/50 ; G01N33/542 ; C12Q1/34 ; G01N33/53 ; C12Q1/68 ; C12N15/09 ; G01N33/58

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 remains undegraded, and the fluorescent dye A remains quenched due to the influence of the fluorescent dye B. By detecting a decrease in the degradation rate of intracellular RNA based on this principle, harmfulness of a chemical substance can be quickly and sensitively evaluated before occurrence of cell death.
Public/Granted literature
- US20180217128A1 METHOD FOR MEASURING IN VIVO INHIBITION OF INTRACELLULAR RNASE Public/Granted day:2018-08-02
Information query