Methods and reagents for molecular proximity detection using RNA-guided nucleic acid binding proteins

Patent metadata

Field	Value
Publication number	US-11268144-B2
Application number	US-201615762748-A
Country	US
Kind code	B2
Filing date	Sep 23, 2016
Priority date	Sep 24, 2015
Publication date	Mar 8, 2022
Grant date	Mar 8, 2022

Abstract

The present disclosure provides reagents and methods for molecular proximity detection of specific endogenous nucleic acids in situ using RNA-guided nucleic acid binding proteins.

First claim

What is claimed is:

1. A complex comprising at least one probe comprising (i) an engineered, non-naturally occurring Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR)-associated (Cas) (CRISPR/Cas) system comprising a Cas9 protein and a guide RNA and (ii) an oligonucleotide that is linked directly or indirectly to the CRISPR/Cas system, wherein the guide RNA comprises the nucleotide sequence of SEQ ID NO:9.

2. The complex of claim 1 , wherein the Cas9 protein comprises two functional nuclease domains, one functional nuclease domain, or no functional nuclease domains.

3. The complex of claim 1 , wherein the complex comprises a first probe comprising (i) the CRISPR/Cas system and (ii) a first oligonucleotide that is linked to the guide RNA by covalent or non-covalent bonds.

Assignees

• Sigma Aldrich Co Llc

Inventors

• Davis Gregory D
• Palhan Vikas B
• Kreader Carol A

Classifications

• C07K16/40 Primary

  against enzymes · CPC title

• C12N2800/80

  Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites · CPC title

• C12Q1/682

  Signal amplification · CPC title

• C12Q1/6804

  Nucleic acid analysis using immunogens (immunoassay G01N33/53) · CPC title

• C12Q1/6853

  using modified primers or templates · CPC title