# US 2014/0196172 A1 · Organelle Targeting Nanocarriers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0196172 A1 - Document ID: US20140196172A1 - Status: Published application - Kind: A1 - Application number: 14/129,386 - Filed: 2012-08-02 - Published: 2014-07-10 - Earliest claimed: 2011-08-04 - Applicants: Her Majesty the Queen in Right of Canada, as Repre sented by the Minister of Agriculture and AgriFoo - Inventors: Francois Eudes; Trevor Macmillan - Web page: https://patentgrep.com/patent/US20140196172A1 ## CPC - C12N 15/8201: {Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation} - C07K 14/415: from plants - C12N 15/8221: {Transit peptides} - C12N 15/85: for animal cells - C07K 2319/07: containing a mitochondrial localisation signal - C07K 2319/08: containing a chloroplast localisation signal - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract Provided are organelle targeting nanocarriers, including peptides, which act to deliver biological molecules such as nucleic acids to non-nuclear organelles such as mitochondria and chloroplasts. Also provided are methods for genetic transformation of non-nuclear organelles using such nanocarriers. ## Claims ### Claim 1 A method for delivering a nucleic acid to a non-nuclear organelle in a cell, the method comprising exposing the cell to a composition comprising at least one nucleic acid and at least one organelle targeting nanocarrier; wherein the at least one nucleic acid translocates across a cell membrane of the cell and enters the non-nuclear organelle in the presence of the at least one organelle targeting nanocarrier. ### Claim 2 (depends on 1) The method according to claim 1 wherein the nucleic acid is DNA. ### Claim 3 (depends on 1) The method according to claim 1 wherein the cell is a plant cell. ### Claim 4 (depends on 1) The method according to claim 1 wherein the cell is an animal cell. ### Claim 5 (depends on 1) The method according to claim 1 wherein the non-nuclear organelle is a mitochondrion. ### Claim 6 (depends on 5) The method according to claim 5 wherein the organelle targeting nanocarrier is a polypeptide having a charge ratio of about 4 to about 7 and hydrophilicity of about 0 to about −0.5. ### Claim 7 (depends on 6) The method according to claim 6 wherein the organelle targeting nanocarrier is a polypeptide having a sequence selected from: MFSYLPRYPLRAASARALVRATRPSYRSALLRYQ; (SEQ ID NO: 1) MAAWMRSLFSPLKKLWIRMH; (SEQ ID NO: 2) MKLLWRLILSRKW; (SEQ ID NO: 3) MWWRRSRTNSLRYT; (SEQ ID NO: 4) and MLFRLRRSVRLRGLLA. (SEQ ID NO: 5) ### Claim 8 (depends on 1) The method according to claim 1 wherein the non-nuclear organelle is a chloroplast. ### Claim 9 (depends on 8) The method according to claim 8 wherein the organelle targeting nanocarrier is a polypeptide having a charge ratio of about 2 to about 4.2 and hydrophilicity of about 0 to about −0.2. ### Claim 10 (depends on 9) The method according to claim 9 wherein the organelle targeting nanocarrier is a polypeptide having a sequence selected from: MGGCVSTPKSCVGAKLR; (SEQ ID NO: 6) MQTLTASSSVSSIQRHRPHPAGRRSSSVTFS; (SEQ ID NO: 7) MKNPPSSFASGFGIR; (SEQ ID NO: 8) MAALIPAIASLPRAQVEKPHPMPVSTRPGLVS; (SEQ ID NO: 9) and MSSPPPLFTSCLPASSPSIRRDSTSGSVTSPLR. (SEQ ID NO: 10) ### Claim 11 A method for producing a genetically modified plant cell, the method comprising: exposing a plant cell containing a non-nuclear organelle to a composition comprising at least one nucleic acid and at least one organelle targeting nanocarrier; wherein the at least one nucleic acid translocates across a cell membrane of the cell and enters the non-nuclear organelle in the presence of the at least one organelle targeting nanocarrier so as to transfect the non-nuclear organelle. ### Claim 12 (depends on 11) The method according to claim 11 wherein the plant cell is an embryogenic microspore. ### Claim 13 (depends on 11) A genetically modified plant cell produced by the method according to claim 11. ### Claim 14 (depends on 11) A method for producing a genetically modified plant, the method comprising: producing a genetically modified plant cell by the method according to claim 11; and generating a plant from the genetically modified plant cell. ### Claim 15 (depends on 14) A genetically modified plant generated by the method according to claim 14. ### Claim 16 A method for producing a genetically modified animal cell, the method comprising: exposing an animal cell containing at least one mitochondrion to a composition comprising at least one nucleic acid and at least one mitochondrion targeting nanocarrier; wherein the at least one nucleic acid translocates across a cell membrane of the cell and enters the at least one mitochondrion in the presence of the at least one mitochondrion targeting nanocarrier so as to transfect the at least one mitochondrion. ### Claim 17 A mitochondrion targeting nanocarrier wherein the mitochondrion targeting nanocarrier is a polypeptide having a mitochondrial targeting peptide sequence, a charge ratio of about 4 to about 7 and a hydrophilicity of about 0 to about −0.5. ### Claim 18 (depends on 17) The mitochondrion targeting nanocarrier according to claim 17 wherein the polypeptide has a sequence selected from: MFSYLPRYPLRAASARALVRATRPSYRSALLRYQ; (SEQ ID NO: 1) MAAWMRSLFSPLKKLWIRMH; (SEQ ID NO: 2) MKLLWRLILSRKW; (SEQ ID NO: 3) MWWRRSRTNSLRYT; (SEQ ID NO: 4) and MLFRLRRSVRLRGLLA. (SEQ ID NO: 5) ### Claim 19 A chloroplast targeting nanocarrier wherein the chloroplast targeting nanocarrier is a polypeptide having a chloroplast transit peptide sequence, a charge ratio of about 2 to about 4.2 and a hydrophilicity of about 0 to about −0.2. ### Claim 20 (depends on 19) The chloroplast targeting nanocarrier according to claim 19 wherein the polypeptide has a sequence selected from: MGGCVSTPKSCVGAKLR; (SEQ ID NO: 6) MQTLTASSSVSSIQRHRPHPAGRRSSSVTFS; (SEQ ID NO: 7) MKNPPSSFASGFGIR; (SEQ ID NO: 8) MAALIPAIASLPRAQVEKPHPMPVSTRPGLVS; (SEQ ID NO: 9) and MSSPPPLFTSCLPASSPSIRRDSTSGSVTSPLR. (SEQ ID NO: 10)