公开(公告)号：US12203080B2

公开(公告)日：2025-01-21

申请号：US17142063

申请日：2021-01-05

Applicant: KEYGENE N.V.

Inventor： Paul Bundock ,  Jeroen Stuurman

IPC: C12N15/82 ,  A01H1/04 ,  C12N9/22

Abstract: The current disclosure relates to the field of plants, in particular to the fields of plant breeding and plant genetics. More particular, the disclosure concerns inventive methodology that may be useful in improving plant properties. In particular the invention may be useful in removing linkage drag. Also provided are plant and plant parts obtained with the method disclosed herein.

公开(公告)号：US20250011800A1

公开(公告)日：2025-01-09

申请号：US18757549

申请日：2024-06-28

Applicant: Keygene N.V.

Inventor： Paul Bundock ,  Michiel Theodoor Jan De Both ,  Franck Georges Paul Lhuissier

IPC: C12N15/82

Abstract: Method for targeted alteration of a duplex acceptor DNA sequence in a plant cell protoplast, comprising combining the duplex acceptor DNA sequence with a donor mutagenic nucleobase, wherein the duplex acceptor DNA sequence contains a first DNA sequence and a second DNA sequence which is the complement of the first DNA sequence and wherein the donor mutagenic nucleobase comprises at least one mismatch with respect to the duplex acceptor DNA sequence to be altered, preferably with respect to the first DNA sequence, wherein the method further comprises a step of introducing the donor mutagenic nucleobase into the cell protoplasts using polyethylene glycol (PEG) mediated transformation and the use of PEG protoplast transformation for enhancing the rate of targeted mutagenesis.

公开(公告)号：US12215329B2

公开(公告)日：2025-02-04

申请号：US16485806

申请日：2018-02-15

Applicant: Keygene N.V.

Inventor： Paul Bundock

IPC: C12N15/82 ,  C12N9/22 ,  C12N9/78 ,  C12N15/11

Abstract: The current invention relates to methods of targeted genetic alteration in cells, preferably plant cells, as well as to plant cells and plants thus obtained using at least a fusion protein comprising a site-specific nuclease domain and a deaminase domain, or a construct encoding the same. The method also provides for a composition and a kit comprising a combination of a first fusion protein comprising a cytosine deaminase domain and a second fusion protein comprising an adenine deaminase domain, preferably for use in the method of the invention. The method provides for targeted alteration of a DNA duplex in plant cells with increased efficacy.

公开(公告)号：US11008579B2

公开(公告)日：2021-05-18

申请号：US15084935

申请日：2016-03-30

Applicant: Keygene N.V.

Inventor： Paul Bundock ,  Michiel Theodoor Jan De Both ,  Frank Lhuissier

IPC: C12N15/82

Abstract: Method for targeted alteration of a duplex acceptor DNA sequence in a plant cell protoplast, comprising combining the duplex acceptor DNA sequence with a donor mutagenic nucleobase, wherein the duplex acceptor DNA sequence contains a first DNA sequence and a second DNA sequence which is the complement of the first DNA sequence and wherein the donor mutagenic nucleobase comprises at least one mismatch with respect to the duplex acceptor DNA sequence to be altered, preferably with respect to the first DNA sequence, wherein the method further comprises a step of introducing the donor mutagenic nucleobase into the cell protoplasts using polyethylene glycol (PEG) mediated transformation and the use of PEG protoplast transformation for enhancing the rate of targeted mutagenesis.

公开(公告)号：US20170166911A1

公开(公告)日：2017-06-15

申请号：US15384644

申请日：2016-12-20

Applicant: Keygene N.V.

Inventor： Paul Bundock ,  Bernarda Gerharda Johanna Fierens-Onstenk ,  Franck Lhuissier

IPC: C12N15/82

CPC classification number: C12N15/8206 ,  C12N15/8201 ,  C12N15/8218 ,  C12N15/8266

Abstract: The invention relates to a method for the introduction of one or more molecules of interest in a plant cell protoplast by providing plant cell protoplasts, performing a first transfection of the plant cell protoplast with a composition that is capable of altering the regulation of one or more pathways selected from the group consisting of Mismatch Repair System and Non-Homologous End Joining and/or a composition that is capable of introducing DSBs, performing a second transfection of the plant cell protoplast with one or more molecules of interest such as mutagenic oligonucleotides and allowing the cell wall to form.

公开(公告)号：US12215363B2

公开(公告)日：2025-02-04

申请号：US16811393

申请日：2020-03-06

Applicant: Keygene N.V.

Inventor： Paul Bundock

IPC: C12N9/22 ,  C12N15/10 ,  C12N15/82 ,  C12N15/90

Abstract: The invention pertains to the targeted alteration of a duplex DNA in a cell, whereby two site-specific nucleases generate an indel, such that the open reading frame is not altered after the second indel. The invention further pertains to the use of such nucleases for the targeted alteration of an open reading frame in duplex DNA and a kit of parts for use in a method of the invention. Using the method of the invention, novel plants were obtained having an improved herbicide resistance. The invention therefore also concerns plants having improved herbicide resistance due to the expression of an altered ALS protein.

公开(公告)号：US11371051B2

公开(公告)日：2022-06-28

申请号：US16310764

申请日：2017-06-20

Applicant: Keygene N.V.

Inventor： Paul Bundock ,  Anita Ketelaars-Bonné

IPC: C12N15/82 ,  C12N9/22 ,  C12N15/11

Abstract: Disclosed is a new method of providing plant cells with a targeted alteration in a DNA molecule. The method comprises contacting a population of plant cells comprising a DNA molecule, the DNA molecule having a target sequence, with an aqueous medium, wherein the aqueous medium comprises a CRISPR associated protein (CAS protein) or a CAS-like protein, and a CRISPR-Cas system guide RNA that hybridizes with the target sequence, and wherein the aqueous medium comprises polyethylene glycol (PEG), but needs to be substantially free of glycerol.

公开(公告)号：US10995338B2

公开(公告)日：2021-05-04

申请号：US14655633

申请日：2013-12-20

Applicant: KEYGENE N.V.

Inventor： Paul Bundock ,  Jeroen Stuurman

IPC: C12N15/82 ,  A01H1/04 ,  C12N9/22

Abstract: The current disclosure relates to the field of plants, in particular to the fields of plant breeding and plant genetics. More particular, the disclosure concerns inventive methodology that may be useful in improving plant properties. In particular the invention may be useful in removing linkage drag. Also provided are plant and plant parts obtained with the method disclosed herein.

公开(公告)号：US09624506B2

公开(公告)日：2017-04-18

申请号：US14640700

申请日：2015-03-06

Applicant: Keygene N.V.

Inventor： Paul Bundock

IPC: C12N15/82 ,  C12N9/10

CPC classification number: C12N15/8275 ,  C12N9/1092 ,  C12Y205/01019

Abstract: The present invention relates to a new method for improving glyphosate resistance of a plant. The method encompasses providing one or more specific mutations in a specific nucleotide sequence in said plant. In comparison to a plant not manipulated according to the method, the plant obtained by the method displays (improved) glyphosate resistance. Also provided are a (transgenic) plant, including a seed thereof, and a plant product that can be obtained by the method according to the invention.