US 2024/0175015 A1 · RELIABLE IDENTIFICATION OF REGIONS ('A SITES') IN COMPLEX RNA MOLECULES THAT ARE ACCESSIBLE TO NUCLEIC ACIDS OR COMPLEXES OF NUCLEIC ACIDS WITH ENDONUCLEASES

Abstract

The invention relates to a method for detecting accessible regions (‘a sites’) in complex RNA molecules (target RNAs), wherein nucleic acids or complexes of these nucleic acids and endonucleases associated therewith bind to the a-sites and alter the function of the target RNAs where by nucleic acids of different types (‘eNAs’) bind to the a-sites and either alone or through an associated endonuclease alter the function of this target RNA. The invention further relates to the use of the method for identifying nucleic acids of different types eNAs that bind to the a-sites and are capable of directing endonucleases, selected from AGO proteins, RNase H and Cas proteins to the a-sites of target RNAs and are capable of, in the presence or absence of these endonucleases reliably affecting/altering the function of these RNA molecules; and to eNAs and a composition containing these eNAs in pathogen control.

Claims17

Dependent claims

Method for the detection of accessible regions (‘a-sites’) in complex RNA molecules (target RNAs), wherein nucleic acids or complexes of these nucleic acids and endonucleases associated therewith bind to the a-sites and alter the function of the target RNAs, the method comprising:

(i) provisioning of a target RNA;
(ii) performing an esiRNA/eRNA screen that identifies siRNAs that can reliably induce a functional change in this target RNA in complexes with Argonaute (AGO) proteins;
(iii) performing a subsequent assay with derived antisense DNA oligonucleotides (ASO) to determine whether they can induce a functional change in the target RNA in the presence or absence of RNase H2 and/or
(iv) performing subsequent testing with g/crRNAs derived therefrom to determine whether they can induce a functional change in the target RNA in the presence of a Cas protein;
and
(v) identifying the a-sites in the target RNA, whereby nucleic acids of different types (‘eNAs’) bind to the a-sites and either alone or through an associated endonuclease alter the function of this target RNA.

3 more elements

Dependencies are inferred from the claim text.

Reliable identification of regions ('a sites') in complex rna molecules that are accessible to nucleic acids or complexes of nucleic acids with endonucleases

Life of this patent

Reliable identification of regions ('a sites') in complex rna molecules that are accessible to nucleic acids or complexes of nucleic acids with endonucleases

Life of this patent

Abstract

Claims17