Methods and compositions for identifying and enriching for cells comprising site specific genomic modifications

Patent metadata

Field	Value
Publication number	US-11898149-B2
Application number	US-202318181704-A
Country	US
Kind code	B2
Filing date	Mar 10, 2023
Priority date	Dec 23, 2014
Publication date	Feb 13, 2024
Grant date	Feb 13, 2024

Abstract

The present invention relates to methods and compositions for modifying a target site in the genome of a plant cell. Such modifications include integration of a transgene and mutations. The present invention also relates to methods and compositions for identifying and enriching for cells which comprise a modified target site.

First claim

What is claimed is:

1. A method of integrating a transgene into a genomic nuclease cleavage site in an event MIR 604 transgenic maize genome, comprising introducing into an event MIR604 maize cell:
a) a first nucleic acid molecule comprising at least 100 contiguous nucleotides, wherein said at least 100 contiguous nucleotides have at least 90% identity with a target site in a nucleotide sequence comprising SEQ ID NO: 135, and further comprising a transgene; and
b) a second nucleic acid molecule comprising a nucleotide sequence encoding a nuclease for site-directed cleavage at a genomic nuclease cleavage site adjacent to a nucleotide sequence with at least 90% identity to a nucleotide sequence comprising SEQ ID NO: 135, that corresponds to the at least 100 contiguous nucleotides of (a),
under conditions wherein expression of the second nucleic acid molecule can occur to produce the nuclease and the nuclease can cleave the nucleotide sequence at the genomic nuclease cleavage site, whereby the transgene is integrated at the genomic nuclease target cleavage site in the event MIR 604 maize genome.

2. A method of producing a maize plant, plant part, or progeny thereof comprising a transgene integrated into a genomic nuclease cleavage site in an event MIR604 maize genome, comprising regenerating a maize plant from the maize cell produced by the method of claim 1 .

3. A maize plant, plant part, or progeny thereof comprising a transgene integrated into a genomic nuclease cleavage site in an event MIR604 maize genome, produced by the method of claim 2 .

Assignees

• Syngenta Participations Ag

Inventors

• Chen Zhongying
• Kim Myoung
• Chilton Mary-Dell
• Zhong Heng
• Gu Weining
• Jiang Yaping
• Que Qiudeng

Classifications

• C12N15/8213 Primary

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

• C12N15/1058

  Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms · CPC title

• C12N15/8205

  Agrobacterium mediated transformation · CPC title

• C12N15/8207

  by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers · CPC title