Method for achieving site specific integration of exogenous DNA delivered by non-biological means to plant cells

• US 6,410,329 B1
• Filed: 05/18/2000
• Issued: 06/25/2002
• Est. Priority Date: 09/25/1995
• Status: Expired due to Term

• Current Assignee: Novartis Finance Corporation (Novartis Ag)
• Original Assignee: Novartis Finance Corporation (Novartis Ag)
• Inventors: Hansen, Genevieve, Suttie, Janet Louise, Chilton, MaryDell
• Primary Examiner(s): Benzion, Gary
• Assistant Examiner(s): Kruse, David H
• Application Number:
  US09/574,132
• Time in Patent Office: 768 Days
• Field of Search: 435/468, 435/470, 435/412, 435/419, 800/278, 800/288, 800/293, 800/320.1
• US Class Current: 435/468
• 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 15/8206 by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers

Abstract

The present invention provides an improved method of site directed integration of an exogenous DNA molecule by co-delivery of a gene of interest flanked by nucleotide sequences recognized by a recombinase and a mRNA encoding the homologous recombinase.

Claims

1. A method of site specific integration of a gene of interest into the genome of a plant cell containing a recombinase site, comprising co-delivery into the plant cell an mRNA encoding a recombinase and a DNA sequence comprising the gene of interest flanked by nucleotide sequences recognized by the recombinase.

2. The method according to claim 1 wherein the recombinase is Cre recombinase.

3. The method of claim 1 wherein the recombinase is FLP recombinase.

4. The method of claim 1 wherein the recombinase is R-recombinase.

5. The method according to claim 1 wherein the means of delivery of the mRNA and the DNA sequence into the plant cell is by microprojectile bombardment.

6. The method according to claim 1 wherein the plant cell is a maize cell.