Method of modifying genome of organism and use thereof

Patent metadata

Field	Value
Publication number	US-2018371477-A1
Application number	US-201616060897-A
Country	US
Kind code	A1
Filing date	Dec 12, 2016
Priority date	Dec 11, 2015
Publication date	Dec 27, 2018
Grant date	—

Abstract

A method of modifying the genome of an organism, wherein the modification method includes modifying the genome of the organism by using in a cell of the organism a protein having an optimal temperature for double-stranded DNA breakage activity in an ordinary temperature region.

First claim

1 . A method of modifying a genome of an organism, wherein the modification method comprises modifying the genome of the organism by using, in a cell of the organism, a protein having an optimal temperature for double-stranded DNA breakage activity in an ordinary temperature region.

2 . The modification method according to claim 1 , wherein the protein is a restriction enzyme originating from a bacterium that is not a thermophile.

3 . The modification method according to claim 1 , wherein the optimal temperature for the double-stranded DNA breakage activity of the protein is at least 25 deg C. and not more than 40 deg C.

4 . The modification method according to claim 1 , wherein the organism is a eukaryotic organism.

5 . The modification method according to claim 1 , wherein the modifying is carried out at at least 20 deg C. and not more than 45 deg C.

6 . The modification method according to claim 5 , wherein the modifying is carried out for at least 10 minutes and not more than 3 hours.

7 . The modification method according to claim 1 that uses the protein obtained by an expression of an exogenous gene that encodes the protein.

8 . The modification method according to claim 1 , wherein the modifying comprises controlling production of the protein to a degree that enables modification of the genome while maintaining growth capacity of the organism.

9 . The modification method according to claim 7 , wherein the modifying comprises modifying the genome of the organism by inducing the expression of the exogenous gene.

10 . The modification method according to claim 7 , wherein the modifying comprises modifying the genome of the organism by continuous maintenance of the expression of the exogenous gene.

11 . The modification method according to claim 1 , wherein the modifying comprises causing an action of the protein by directly supplying the protein into the cell of the organism.

12 . The modification method according to claim 1 , wherein the protein is one or more restriction enzymes selected from the group consisting of HinP1I, MseI, HaeIII, AluI, MboI, and HbaI.

13 . The modification method according to claim 1 , wherein the organism is a plant body or a portion of a plant body.

14 . The modification method according to claim 13 , wherein the plant body or the portion of the plant body is one or more selected the group consisting of a seed, a shoot apex, a lateral bud, a flower bud, pollen, an ovary, an endosperm, and an embryo, and a portion of a seed, a shoot apex, a lateral bud, a flower bud, pollen, an ovary, an endosperm, and an embryo.

15 . The modification method according to claim 1 , wherein the organism is a microorganism.

16 . A method of producing a population of genomically modified organisms, wherein the production method comprises modifying a genome of a parent organism by causing an action, within a cell of the parent organism, of a protein having an optimal temperature for double-stranded DNA breakage activity in an ordinary temperature region.

17 . A method of producing a genomically modified organism, wherein the production method comprises:
modifying a genome of a parent organism by causing an action, in a cell of the parent organism, of a protein having an optimal temperature for double-stranded DNA breakage activity in an ordinary temperature region; and selecting an intended eukaryotic organism from a population of eukaryotic organisms that carry modified genomes based on an indicator.

18 . A breeding material, wherein the breeding material comprises DNA that has a coding region that encodes a protein that has a double-stranded DNA breakage activity that exhibits an optimal temperature in an ordinary temperature region wherein the coding region contains an intron that is not processed within a host for the breeding material.

19 . The breeding material according to claim 18 , which is an expression vector for a eukaryotic organism.

20 . The breeding material according to claim 18 , wherein the host is a prokaryotic organism.

Assignees

• Toyota Chuo Kenkyusho Kk
• Univ Tokyo
• Univ Meiji

Inventors

• Tanaka Hidenori
• Muramoto Nobuhiko
• Sugimoto Hiroki
• Mitsukawa Norihiro
• Ohto Chikara
• Kondo Satoshi
• Ohta Kunihiro
• Nakamura Takahiro
• Osato Shuichi

Classifications

• C12N15/09

  Recombinant DNA-technology · CPC title

• C12N15/102

  Mutagenizing nucleic acids · CPC title

• C12N15/8206

  by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated · CPC title

• C12N15/01

  Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor · CPC title

• C12N15/8213 Primary

  Targeted insertion of genes into the plant genome by homologous recombination · CPC title