Methods for altering the genome of a monocot plant cell

• US 8,912,392 B2
• Filed: 06/27/2008
• Issued: 12/16/2014
• Est. Priority Date: 06/29/2007
• Status: Active Grant

• Current Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Original Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Inventors: Lyznik, L. Aleksander, Tao, Yumin, Gao, Huirong
• Primary Examiner(s): Fox, David T
• Assistant Examiner(s): KEOGH, MATTHEW R
• Application Number:
  US12/147,834
• Publication Number:
  US 20090133152A1
• Time in Patent Office: 2,363 Days
• Field of Search: None
• US Class Current: 800/278
• CPC Class Codes: 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
• CPC Class Codes: C12N 15/8213 Targeted insertion of genes into the plant genome by homologous recombination
• CPC Class Codes: C12N 15/8241 Phenotypically and genetically modified plants via recombinant DNA technology
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/13 Plant traits
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

Methods and compositions for altering the genome of a monocot plant cell, and a monocot plant are disclosed. The methods and compositions use a double-strand break inducing agent to alter a monocot plant or plant cell genomic sequence comprising a recognition sequence for the double-strand break inducing agent.

Claims

1. A method of modifying a specific endogenous genomic target sequence comprising:

(a) contacting at least one monocot plant cell comprising the endogenous genomic target sequence in its genome with (i) a phenotypic marker, (ii) a DNA fragment, and (iii) an engineered double-strand break inducing agent capable of inducing a double-strand break at the endogenous genomic target sequence, wherein the engineered double-strand break inducing agent comprises SEQ ID NO;

21;

(b) selecting cells comprising the phenotypic marker wherein the phenotypic marker is not indicative of integration at the endogenous genomic target sequence; and

(c) identifying cells from step (b) having an alteration in their genome at the endogenous genomic target sequence wherein the alteration is selected from the group consisting of (i) replacement of at least one nucleotide, (ii) a deletion of at least one nucleotide, (iii) an insertion of at least one nucleotide, and (iv) any combination of (i)-(iii),

wherein identifying comprises any method that does not use the phenotypic marker.

2. The method of claim 1, wherein the monocot plant cell is from maize, rice, sorghum, barley, wheat, millet, oats, sugarcane, turfgrass, or switch grass.

3. The method of claim 1, wherein the monocot plant cell is from maize.

4. The method of claim 1, further comprising recovering a fertile monocot plant having the alteration in its genome.