Processing

Please wait...

Settings

Settings

Goto Application

1. WO2020117553 - GENE SILENCING VIA GENOME EDITING

Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

[ EN ]

What is claimed is:

1) A method of reducing expression a target gene comprised of:

a) Introducing into a cell a nuclease capable of site -directed DNA cleavage at a target genomic site;

b) Making two or more double strand cuts within a single target gene;

c) Selecting for a cell where the double strand cuts have been repaired with the intervening DNA inverted;

d) reducing expression of the target gene.

2) The method of claim 1 , wherein the nuclease is selected from the group consisting of

meganucleases (MNs), zinc-finger nucleases (ZFNs), transcription-activator like effector nucleases (TALENs), Cas9 nuclease, Cfpl nuclease, dCas9-FokI, dCpfl-Fokl, chimeric

Cas9/Cpfl-cytidine deaminase, chimeric Cas9/Cpfl -adenine deaminase, chimeric FENl-FokI, and Mega-TALs, a nickase Cas9 (nCas9), chimeric dCas9 non-Fokl nuclease and dCpfl non- Fokl nuclease.

3) The method of claim 1 wherein the double strand cuts in the target gene are located in the

promoter, UTR, exon, intron, or gene-gene junction region.

4) The method of claim 1 wherein the cell of claim 1 has a haploid, diploid, polyploid, or hexiploid genome.

5) The method of claiml wherein the target gene is recessive or semi-dominant.

6) The method of claim 1 further comprising one or more guide sequences.

7) The method of claim 6 wherein the one or more guide sequences comprise two or more guide sequences.

8) The method of claim 1 wherein the cell is a plant cell.

9) A method of rearranging a chromosome by genome editing, comprising:

a. generating at least one breakage in the chromosome by a site-directed nuclease;

b. selecting a chromosome with a rearrangement.

10) The method of claim 9, wherein the site -directed nuclease is selected from the group consisting of meganucleases (MNs), zinc-finger nucleases (ZFNs), transcription-activator like effector nucleases (TALENs), Cas9 nuclease, Cfpl nuclease, dCas9-FokI, dCpfl-Fokl, chimeric

Cas9/Cpfl-cytidine deaminase, chimeric Cas9/Cpfl -adenine deaminase, chimeric FENl-FokI, and Mega-TALs, a nickase Cas9 (nCas9), chimeric dCas9 non-Fokl nuclease and dCpfl non- Fokl nuclease.

11) The method of claim 9, wherein the chromosome rearrangement comprises a deletion, duplication, inversion, or translocation.

12) The method of claim 9, wherein the chromosome rearrangement causes a modification of gene expression.

13) The method of claim 9, wherein the gene expression modification includes regulation at precursor mRNA level, or at mature mRNA level or at translation level.

14) The method of claim 9, wherein the chromosome rearrangement includes chromosomes from two species when the chromosomes can be grouped in one nuclei such as in an interspecific hybrid.

15) The method of claim 9, wherein the chromosome rearrangement leads to new allele generation via fusing at least two alleles or two components from different alleles.

16) The method of claim 11, wherein chromosome rearrangement is targeted to a promoter, exon, intron, or transcription terminator.

17) The method of claim 12, chromosome rearrangement causes a modification of gene expression of different genes with sequence similarity to the rearranged gene.

18) The method of claim 11 , wherein the deletion, duplication, inversion, or translocation is no less than 19 base pairs.