# US 10,767,176 B2 · CRISPR-based compositions and methods of use

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- Number: US 10,767,176 B2
- Document ID: US10767176B2
- Status: Granted
- Kind: B2
- Application number: 15/299,549
- Filed: 2016-10-21
- Published: 2017-02-16
- Granted: 2020-09-08
- Earliest claimed: 2014-12-18
- Assignees: Integrated Dna Technologies, Inc.
- Applicants: Integrated Dna Technologies, Inc.
- Inventors: Michael Allen Collingwood; Ashley Mae Jacobi; Garrett Richard Rettig; Mollie Sue Schubert; Mark Aaron Behlke
- Web page: https://patentgrep.com/patent/US10767176B2

## CPC

- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12N 9/222: {Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes}
- C12N 2310/10: Type of nucleic acid
- C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR]
- C12N 2310/315: Phosphorothioates
- C12N 2310/317: with an inverted bond, e.g. a cap structure
- C12N 2310/321: 2'-O-R Modification
- C12N 2310/322: 2'-R Modification
- C12N 2310/3231: having an additional ring, e.g. LNA, ENA
- C12N 2310/346: having a combination of backbone and sugar modifications
- C12N 2310/531: Stem-loop; Hairpin
- C12N 2320/51: modulating the chemical stability, e.g. nuclease-resistance
- C12N 2320/53: reducing unwanted side-effects
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

This invention pertains to modified compositions for use in CRISPR systems, and their methods of use. In particular, length-modified and chemically-modified forms of crRNA and tracrRNA are described for use as a reconstituted guide RNA for interaction with Cas9 of CRISPR systems. The resultant length-modified and chemically-modified forms of crRNA and tracrRNA are economical to produce and can be tailored to have unique properties relevant to their biochemical and biological activity in the context of the CRISPR Cas9 endonuclease system.

## Claims

### Claim 1

An isolated tracrRNA selected from the group consisting of SEQ ID NOS:19-21, and 449-451, wherein the isolated tracrRNA is active in a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein endonuclease system.