US 10,851,420 B2 · Editing profiling of PDE8A pre-mRNA: use as specific biomarker of ADARs activities in human tissues to diagnose and to predict and assess therapeutic efficacy and/or efficiency or potential drug side effects

Abstract

The present invention relates to the use of the editing profile of PDE8A pre-mRNA as a specific bio marker of ADARs activities in evolved primate, particularly in Human tissues. The present invention also relates to an in vitro method for predicting in Human an alteration of the mechanism of the ADARs catalysed pre-mRNA editing of target genes, by analysing the PDE8A pre-mRNA editing profile in a peripheral tissue sample containing cells expressing said PDE8A pre-mRNA, such as blood sample. The present invention is also directed to an in vitro method for the screening of potential therapeutic compound and to predict and assess therapeutic efficacy and/or efficiency or to diagnose potential severe brain or peripheral drug side effects implementing said PDE8A pre-mRNA editing profile as specific biomarker. The present invention is further directed to a method for determining the PDE8A pre-mRNA editing profile in Human, particularly by capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) method after amplification by a nested PCR. Finally the invention relates to particular nucleic acid primers implemented in said nested PCR and kit comprising such sets of primers and human cells capable of expressing PDE8A and ADARs.

Claims16

Dependent claims

An in vitro method, comprising:

a) providing a biological subject sample comprising Peripheral Blood Mononuclear Cells (PBMC), said PBMC expressing the editing enzymes ADAR1a, ADAR1b and ADAR2, and the phosphodiesterase subtype 8A (PDE8A);
b) preparing a cellular RNA extract from the subject sample;
and
c) determining the editing profile of the PDE8A pre-mRNA in said cellular RNA extract, wherein determining the editing profile of the PDE8A pre-mRNA comprises detecting the ned (non edited isoform) and B isoforms.

1 more element

Dependencies are inferred from the claim text.

Facts

Application

16/442,825Filed 17 Jun 2019 · division

Priority

24 Jun 2010

Published

US 2020/0032338 A130 Jan 2020

Granted

1 Dec 2020

Term ends

Not estimatedThe source records don’t support an estimate.

Classification17 codes

C12Q 1/6883
Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes: for diseases caused by alterations of genetic material
C12Q 1/34
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
G01N 33/5023
For testing non-proliferative effects: on expression patterns
C12Q 2600/106
Oligonucleotides characterized by their use: pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism

Family

Application61/344,298Us provisional application · filed 24 Jun 2010

PCT/EP2011/060644Parent of this continuation · filed 24 Jun 2011

Application13/959,928Parent of this continuation · filed 6 Aug 2013

Application15/479,648Parent of this division · filed 5 Apr 2017

US 2020/0032338 A1Pre-grant publication · published 30 Jan 2020

US 10,323,283 B2Parent of this division

Application15/479,648Parent of this continuation

Application13/805,902Parent of this continuation

US 2015/0056622 A1Published application

US 2018/0010184 A1Published application

US 10,851,420 B2This patent

From the US filing history. Foreign family members aren’t listed.

Editing profiling of PDE8A pre-mRNA: use as specific biomarker of ADARs activities in human tissues to diagnose and to predict and assess therapeutic efficacy and/or efficiency or potential drug side effects

Life of this patent

Editing profiling of PDE8A pre-mRNA: use as specific biomarker of ADARs activities in human tissues to diagnose and to predict and assess therapeutic efficacy and/or efficiency or potential drug side effects

Life of this patent

Abstract

Claims16