EP0480971A1 - A method for introducing molecules, particularly genetic material, into plant cells - Google Patents
A method for introducing molecules, particularly genetic material, into plant cellsInfo
- Publication number
- EP0480971A1 EP0480971A1 EP19900910596 EP90910596A EP0480971A1 EP 0480971 A1 EP0480971 A1 EP 0480971A1 EP 19900910596 EP19900910596 EP 19900910596 EP 90910596 A EP90910596 A EP 90910596A EP 0480971 A1 EP0480971 A1 EP 0480971A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- plant cells
- cells
- molecules
- genetic material
- ultrasound treatment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8206—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
Definitions
- the method according to the invention is a useful alternative to previously known introduction methods.
- the method provides a new approach involving only moderate costs, said method being carried out quickly and simply. Based on introduction experiments already conducted it can be predicted that the method will also turn out to be superior when used for a great number of other biological materials, where known methods are insufficient.
- the plant cells are subjected to an ultrasound treatment considerably milder than the ultrasound treatment used for homogenizing or lysing cells. It is important that the treatment is suitably mild so that a sufficient number of plant cells remain viable.
- the plant cell suspension can advantageously be exposed to ultrasonic waves of a frequency range of from 5 kHz to 10 MHz, particularly from 10 to 100 kHz, and an electric output power (i.e.
- the method according to the invention has advantageously been used to introduce molecules into cells of sugar beet and tobacco plants.
- the medium can be any suitable, conventional medium for cells or protoplasts.
- the method according to the invention uses a technique presumbably resulting in a temporary moderate weakening of both the cell membrane and the cell wall, which is shown in intact plant cells in the following examples. The examples thus show that plasmid DNA can penetrate plant cells having been treated with ultrasound.
- CPW is an aqueous solution of a mixture of inorganic salts comprising i.a. approx. 10 mM Ca , described by Frearson et al., (Dev. Biol., 33, 130-137, 1973).
- a cell suspension for ultrasound treatment is prepared by removing cells 3 to 4 days after sub-culturing and washing them twice with CPW 13S (i.e. CPW containing 13% sorbitol), finally suspending said cells in CPW 13S at a ratio of 1 part by volume cells to 4 parts by volume CPW 13S. Then plasmid DNA is added to the suspension in 0.35 ml CPW containing 21% sucrose and plant cells (500.000/ml) in an Eppendorf tube, the final plasmid concentration being 45 ⁇ g/ml.
- CPW 13S i.e. CPW containing 13% sorbitol
- the plasmid DNA used is a plasmid coding for the marker enzyme chloramphenicol-acetyltransferase (CAT) , in this case the plasmid pCaMVCN having the code 27-4909, available from Pharmacia LKB Biotechnology, Uppsala, Sweden.
- CAT chloramphenicol-acetyltransferase
- the present example illustrates the introduction of plasmid DNA into intact sugar beet cells of the genotype Ml.
- the suspension culture of sugar beet cells is maintained by sub-culturing as described above and is treated with ultrasound, cultured and analysed in the manner described.
- the results of the measured CAT-activity appear from Table 1.
- the present example illustrates the introduction of plasmid DNA into intact tobacco cells.
- the cells are removed 3 to 4 days after sub-culturing and washed twice with CPW 13S (i.e. CPW containing 13% sorbitol), finally suspending said cells in CPW 13S at a ratio of 1 part by volume cells to 4 parts by volume CPW 13S.
- Samples of 0.35 ml each are taken out and the plasmid pCaMVCN is added to each sample, the final plasmid concentration being 100 ⁇ g/ml.
- the cells are then subjected to ultrasound treatment under the conditions appearing from Table 2. After culturing for 2 days in the above-mentioned medium the cells are extracted and their CAT-activity is measured. The results appear from Table 2.
- the method according to the invention can be used to introduce molecules into intact plant cells.
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Procédé d'introduction de molécules, notamment de matière génétique dans des cellules végétales intactes, consistant à soumettre un milieu contenant lesdites céllules végétales ainsi que lesdites molécules à un traitement ultrasonique doux.Method for introducing molecules, in particular genetic material, into intact plant cells, comprising subjecting a medium containing said plant cells as well as said molecules to a mild ultrasonic treatment.
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK3251/89 | 1989-06-29 | ||
DK325189A DK168302B1 (en) | 1989-06-29 | 1989-06-29 | Method of introducing molecules, especially genetic material into plant cells |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0480971A1 true EP0480971A1 (en) | 1992-04-22 |
Family
ID=8120791
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP19900910596 Ceased EP0480971A1 (en) | 1989-06-29 | 1990-06-28 | A method for introducing molecules, particularly genetic material, into plant cells |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP0480971A1 (en) |
JP (1) | JPH05500304A (en) |
AU (1) | AU645260B2 (en) |
DK (1) | DK168302B1 (en) |
WO (1) | WO1991000358A1 (en) |
Families Citing this family (38)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2042093C (en) * | 1990-05-09 | 2002-12-24 | Gyula Hadlaczky | Cell line carrying an excess of mammalian centromeres |
CA2231400A1 (en) | 1995-09-08 | 1997-03-13 | Genentech, Inc. | Vegf-related protein |
US6030945A (en) * | 1996-01-09 | 2000-02-29 | Genentech, Inc. | Apo-2 ligand |
US6998116B1 (en) | 1996-01-09 | 2006-02-14 | Genentech, Inc. | Apo-2 ligand |
US5693512A (en) * | 1996-03-01 | 1997-12-02 | The Ohio State Research Foundation | Method for transforming plant tissue by sonication |
US6469144B1 (en) | 1996-04-01 | 2002-10-22 | Genentech, Inc. | Apo-2LI and Apo-3 polypeptides |
US6025155A (en) * | 1996-04-10 | 2000-02-15 | Chromos Molecular Systems, Inc. | Artificial chromosomes, uses thereof and methods for preparing artificial chromosomes |
US6077697A (en) * | 1996-04-10 | 2000-06-20 | Chromos Molecular Systems, Inc. | Artificial chromosomes, uses thereof and methods for preparing artificial chromosomes |
US5851984A (en) * | 1996-08-16 | 1998-12-22 | Genentech, Inc. | Method of enhancing proliferation or differentiation of hematopoietic stem cells using Wnt polypeptides |
US6159462A (en) * | 1996-08-16 | 2000-12-12 | Genentech, Inc. | Uses of Wnt polypeptides |
US6462176B1 (en) | 1996-09-23 | 2002-10-08 | Genentech, Inc. | Apo-3 polypeptide |
US5990281A (en) * | 1996-09-30 | 1999-11-23 | Genentech, Inc. | Vertebrate smoothened proteins |
US6342369B1 (en) | 1997-05-15 | 2002-01-29 | Genentech, Inc. | Apo-2-receptor |
AU739350B2 (en) | 1997-06-05 | 2001-10-11 | University Of Texas System, The | APAF-1, the CED-4 human homolog, an activator of caspase-3 |
EP2083079A1 (en) | 1997-06-18 | 2009-07-29 | Genentech, Inc. | Apo-2DcR |
US6342220B1 (en) | 1997-08-25 | 2002-01-29 | Genentech, Inc. | Agonist antibodies |
EP1045906B1 (en) | 1998-01-15 | 2008-10-15 | Genentech, Inc. | Apo-2 ligand |
US6114603A (en) * | 1998-03-27 | 2000-09-05 | John Innes Center | Genetic engineering of sugarbeet plants |
DE19834612A1 (en) * | 1998-07-31 | 2000-02-24 | Dornier Medtech Holding Int Gmbh | Method for intracellular transfer of oligonucleotides and device for carrying out the same |
US6195936B1 (en) * | 1999-02-22 | 2001-03-06 | University Of Iowa Research Foundation | Method for uptake of a substance into a seed |
MXPA01013236A (en) | 1999-06-28 | 2002-06-21 | Genentech Inc | Methods for making apo-2 ligand using divalent metal ions. |
DE19962904A1 (en) * | 1999-12-23 | 2001-08-09 | Dornier Medizintechnik | Apparatus continuously introducing molecules into cells, avoiding their total disruption, focuses controlled acoustic pulses at axis of duct passing carrier fluid |
ES2267593T3 (en) | 2000-02-16 | 2007-03-16 | Genentech, Inc. | ANTI-APRIL ANTIBODIES AND HYBRIDOMA CELLS. |
EP2390256A1 (en) | 2001-05-30 | 2011-11-30 | Agrisoma, Inc. | Plant artificial chromosomes, uses thereof and methods of preparing plant artificial chromosomes |
NZ530985A (en) | 2001-08-29 | 2006-03-31 | Genentech Inc | Bv8 nucleic acids and polypeptides with mitogenic activity |
WO2003029420A2 (en) | 2001-10-02 | 2003-04-10 | Genentech, Inc. | Apo-2 ligand variants and uses thereof |
ATE482716T1 (en) | 2001-11-13 | 2010-10-15 | Genentech Inc | COMPOSITIONS BASED ON APO2 LIGAND/TRAIL AND THEIR USE |
DE10223196B4 (en) | 2002-05-24 | 2004-05-13 | Dornier Medtech Systems Gmbh | Method and device for transferring molecules into cells |
AU2003247609A1 (en) | 2002-06-24 | 2004-01-06 | Genentech, Inc. | Apo-2 ligand/trail variants and uses thereof |
EP2526960A1 (en) | 2003-03-12 | 2012-11-28 | Genentech, Inc. | Use of BV8 and/or EG-VEGF to promote hematopoiesis |
PT1631313E (en) | 2003-06-05 | 2015-07-02 | Genentech Inc | Combination therapy for b cell disorders |
EP1671627B8 (en) | 2004-12-15 | 2010-04-07 | Dornier MedTech Systems GmbH | Improvement of cell therapy and tissue regeneration in patients with cardiovascular and neurological diseases by means of shockwaves |
GB2452543B (en) * | 2007-09-07 | 2012-07-25 | Wei Huang | Nucleic acid transfer techniques |
ES2565202T3 (en) | 2007-10-16 | 2016-04-01 | Zymogenetics, Inc. | Combination of transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) and anti-CD20 agents for the treatment of autoimmune diseases |
US8669085B2 (en) | 2009-02-05 | 2014-03-11 | Ut-Battelle, Llc | Transformation of gram positive bacteria by sonoporation |
EP3431588A1 (en) | 2009-08-11 | 2019-01-23 | F. Hoffmann-La Roche AG | Production of proteins in glutamine-free cell culture media |
CA2834776A1 (en) | 2011-05-03 | 2012-11-08 | Genentech, Inc. | Therapeutic apo2l/trail polypeptides and death receptor agonist antibodies |
KR101791563B1 (en) * | 2015-05-11 | 2017-11-02 | 대한민국 | Method for delaying the ripening of fruits using sound treatment |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3537261A1 (en) * | 1985-10-19 | 1987-04-30 | Gca Corp | METHOD AND MEDIUM FOR FIELD-INDUCED INSERTING MACROMOLECULES IN LIVING CELLS |
AU615905B2 (en) * | 1987-05-05 | 1991-10-17 | Novartis Ag | Plant tissue transformation |
GB8721015D0 (en) * | 1987-09-07 | 1987-10-14 | Amersham Int Plc | Modifying living cells |
-
1989
- 1989-06-29 DK DK325189A patent/DK168302B1/en not_active IP Right Cessation
-
1990
- 1990-06-28 WO PCT/DK1990/000166 patent/WO1991000358A1/en not_active Application Discontinuation
- 1990-06-28 EP EP19900910596 patent/EP0480971A1/en not_active Ceased
- 1990-06-28 JP JP51026590A patent/JPH05500304A/en active Pending
- 1990-06-28 AU AU60432/90A patent/AU645260B2/en not_active Expired - Fee Related
Non-Patent Citations (1)
Title |
---|
See references of WO9100358A1 * |
Also Published As
Publication number | Publication date |
---|---|
DK325189D0 (en) | 1989-06-29 |
JPH05500304A (en) | 1993-01-28 |
WO1991000358A1 (en) | 1991-01-10 |
DK168302B1 (en) | 1994-03-07 |
DK325189A (en) | 1990-12-30 |
AU6043290A (en) | 1991-01-17 |
AU645260B2 (en) | 1994-01-13 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 19911231 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE DK ES FR GB IT LI LU NL SE |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: SANDOZ LTD. Owner name: SANDOZ-ERFINDUNGEN VERWALTUNGSGESELLSCHAFT M.B.H. Owner name: SANDOZ-PATENT-GMBH |
|
17Q | First examination report despatched |
Effective date: 19931213 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED |
|
18R | Application refused |
Effective date: 19940426 |