US 8,012,752 B2 · In vitro methods for the induction and maintenance of plant cell lines as single suspension cells with intact cell walls, and transformation thereof

Abstract

The subject invention provides simple and consistent methods to break suspension cell aggregates to single cells with intact primary cell walls. The subject invention relates in part to cell separation of suspension cell aggregates cultured in medium containing pectin-degrading enzymes or tubulin de-polymerizing compounds including colchicine. The subject invention also relates to novel uses of compounds for such purposes. Another aspect of the subject invention relates to transformation of the subject, isolated cells. Such processes simplify and integrate single-cell-based transformation and selection processes into transgenic and transplastomic event-generation work processes. The subject invention also removes technical constraints and produces marker-free and uniformly expressing transgenic lines in a high throughput fashion to support various needs of animal health, biopharma, and trait and crop protection platforms.

Claims3

Dependent claims

A method for producing a single plant cell, wherein said method comprises culturing plant cells comprising intact cell walls in medium containing glycerol and a separating agent selected from the group consisting of a pectin-degrading enzyme and a tubulin de-polymerizing compound, wherein said tubulin de-polymerizing compound conforms to the following formula:

wherein
X=, OCH2CO2R,
Y=Cl,
Ar1=unsubstituted phenyl,
Ar2−unsubstituted phenyl;
and
R=2carbon linear.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application

11/965,543Filed 27 Dec 2007 · us provisional application

Priority

29 Dec 2006

Published

US 2010/0159598 A124 Jun 2010

Granted

6 Sep 2011

Term ends

Not estimatedThe source records don’t support an estimate.

Classification4 codes

C12N 5/04
Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor: plant cells…
C12N 5/0025
Culture media for cell or tissue culture: culture media for plant cell or plant tissue culture
C12N 15/8201
For plant cells , e.g. plant artificial chromosomes: methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient…
C12N
Microorganisms or enzymes

In vitro methods for the induction and maintenance of plant cell lines as single suspension cells with intact cell walls, and transformation thereof

Life of this patent

In vitro methods for the induction and maintenance of plant cell lines as single suspension cells with intact cell walls, and transformation thereof

Life of this patent

Abstract

Claims3