# US 10,308,935 B2 · Methods for triggering de novo formation of heterochromatin and or epigenetic silencing with small RNAS

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,308,935 B2
- Document ID: US10308935B2
- Status: Granted
- Kind: B2
- Application number: 15/321,159
- Filed: 2015-06-22
- Published: 2017-06-01
- Granted: 2019-06-04
- Earliest claimed: 2014-06-23
- Assignees: Friedrich Miescher Institute for Biomedical Research
- Applicants: Marc Buehler; Katarzyna Kowalik; Yukiko Shimada
- Inventors: Marc Buehler; Katarzyna Kowalik; Yukiko Shimada
- Web page: https://patentgrep.com/patent/US10308935B2

## CPC

- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- A61K 31/7105: Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12N 2310/14: interfering nucleic acids [NA]
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

The present application relates to a method for the targeted formation of heterochromatin and/or induction of epigenetic gene silencing in a cell using a small RNA, said method comprising the step of inhibiting the Paf1 complex in said cell and the step of contacting said cell with a small RNA targeted to a region of the genome of the cell, said region being the region where heterochromatin formation and/or induction of epigenetic gene silencing should be induced.

## Claims

### Claim 1

A method for the targeted formation of heterochromatin and epigenetic silencing at a site in the genome of an animal cell, wherein the site encodes an expressed RNA,
said method comprising inhibiting the Paf1 complex in the cell and contacting the cell with a small RNA that is sufficiently complementary to a 5′-region of the expressed RNA to hybridize to it under physiological conditions,
wherein the method induces heterochromatin formation and epigenetic gene silencing in said site, and wherein the expressed RNA does not encode any component of the Paf1 complex.

### Claim 2 (depends on 1)

The method of claim 1, wherein the step of inhibiting the Paf1 complex comprises the step of inhibiting a component of the Paf1 complex selected from Paf1, Cdc73, Rtf1, Leo1, and Ctr9.

### Claim 3 (depends on 1)

The method of claim 1, wherein the step of inhibiting the Paf1 complex comprises the step of selecting a mutant cell in which at least one gene of a component of the Paf1 complex is mutated in such a way that the expression of the at least one gene of a component of the Paf1 complex is reduced.

### Claim 4 (depends on 3)

The method of claim 3 wherein said mutant is a naturally-occurring mutant.

### Claim 5 (depends on 2)

The method of claim 2, wherein said component of the Paf1 complex is Leo 1.

### Claim 6 (depends on 2)

The method of claim 2, wherein the expression of said component of the Paf1 complex is reduced.

### Claim 7 (depends on 3)

The method of claim 3, wherein said component of the Paf1 complex is selected from Paf1, Cdc73, Rtf1, Leo1, and Ctr9.

### Claim 8 (depends on 1)

The method of claim 1, wherein said method is carried out in vitro, in vivo or ex vivo.