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WO2000008195A1 - Procede et dispositif de transfert d'oligonucleotides dans des cellules - Google Patents

Procede et dispositif de transfert d'oligonucleotides dans des cellules Download PDF

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Publication number
WO2000008195A1
WO2000008195A1 PCT/DE1999/002327 DE9902327W WO0008195A1 WO 2000008195 A1 WO2000008195 A1 WO 2000008195A1 DE 9902327 W DE9902327 W DE 9902327W WO 0008195 A1 WO0008195 A1 WO 0008195A1
Authority
WO
WIPO (PCT)
Prior art keywords
oligonucleotides
transferred
cell
cells
shock wave
Prior art date
Application number
PCT/DE1999/002327
Other languages
German (de)
English (en)
Inventor
Stefan Endres
Michael Delius
Friedrich Ueberle
Original Assignee
Dornier Medtech Holding International Gmbh
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Dornier Medtech Holding International Gmbh filed Critical Dornier Medtech Holding International Gmbh
Priority to AU63241/99A priority Critical patent/AU6324199A/en
Publication of WO2000008195A1 publication Critical patent/WO2000008195A1/fr

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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
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Definitions

  • the invention relates to a method for intracellular transfer of oligonucleotides according to method claim 1 and a device for carrying out the method according to device claim 4
  • oligonucleotides The intracellular transfer of oligonucleotides is used to specifically inhibit the synthesis of individual proteins in the cell.
  • a short-chain synthetic nucleic acid with a freely selectable sequence of bases is introduced into the cytoplasm of the cell as soon as the cell nucleus of the cell is used to synthesize a If the so-called mRNA, which is necessary for the protein, is released, the oligonucleotide in the cell can take the place of the mRNA, the sequence of which is complementary to the base sequence of the gonucleotide.
  • This “application” blocks the synthesis of exactly one protein. This cell is therefore absent an otherwise present protein The absence of this protein can result in a change in the cell's properties or mortality
  • carrier lipids are used, by means of which an improvement in the uptake of the gonucleotides into the cytoplasm of the cell is produced.
  • a disadvantage of this known method is the need for these relatively expensive carrier lipids in addition to the Having to apply oligonucleotides as consumable for intracellular transfer
  • Another known possibility of transferring oligonucleotides provides for direct introduction, for example by means of microinjections or electroporation.
  • direct introduction for example by means of microinjections or electroporation.
  • the effectiveness of the method of direct introduction is disadvantageous
  • cavitation occurs in liquids under the influence of shock waves.
  • This cavitation can be represented simply as the formation and movement of bubbles or cavities in a liquid.
  • a very fine, needle-like liquid jet is formed. This liquid jet penetrates the cell membrane and transfers a small amount of the liquid into the cell.
  • Oligonucleotides there is a probability, depending on the concentration of the oligonucleotides in the liquid, that oligonucleotides have been transferred into the cell.
  • FIG. 1 shows a schematic structure of a device 1 with which the method according to the invention can be carried out.
  • the device 1 is filled with a liquid 2.
  • the liquid 2 can be exposed to a shock wave by means of a shock wave generator 3.
  • a sample container 4 is partially immersed in the liquid 2.
  • the sample container 4 is attached to a holder, not shown for reasons of clarity.
  • This solution 5 contains the oligonucleotides and the target cells into which the oligonucleotides are to be transferred.
  • the material of the sample container 4 is continuous for shock waves, the essentially aqueous solution 5 in which the oligonucleotides and the target cells for the transfer of the oligonucleotides also pass on the shock waves.
  • Fluid jet hits the cell, it penetrates the cell membrane like a microinjection. A small amount of the liquid that makes up the fluid jet remains in the cell.
  • the oligonucleotides to be transferred are stochastically distributed in the liquid, the oligonucleotides are also transferred stochastically to individual cells.
  • Activating the shock wave source several times increases the probability that oligonucleotides will be transferred into the cell.
  • the target cells can be in a living being, and the oligonucleotides can be located near the target cells by suitable measures (such as blood circulation or injection) Extracorporeal shock waves in the body of the living being and focusing on the one area of the body in which the target cells are located, it is possible to introduce locally limited oligonucleotides into cells of the body and thus change the properties of the cells and / or their mortality or lethality.
  • suitable measures such as blood circulation or injection
  • shock wave source for the purpose of transferring oligonucleotides in cells both in vivo and in vitro represents a method in which harmful influences on the cells, for example through the currents during electroporation or side effects caused by the carrier lipids, are advantageously avoided.

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Plant Pathology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Mechanical Engineering (AREA)
  • Cell Biology (AREA)
  • Sustainable Development (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Des oligonucléotides sont transférés dans des cellules par l'action d'ondes de choc.
PCT/DE1999/002327 1998-07-31 1999-07-30 Procede et dispositif de transfert d'oligonucleotides dans des cellules WO2000008195A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU63241/99A AU6324199A (en) 1998-07-31 1999-07-30 Method and device for the transfer of oligonucleotides in cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19834612A DE19834612A1 (de) 1998-07-31 1998-07-31 Verfahren zum intrazellulären Transfer von Oligonukleotiden und Vorrichtung zur Durchführung desselben
DE19834612.3 1998-07-31

Publications (1)

Publication Number Publication Date
WO2000008195A1 true WO2000008195A1 (fr) 2000-02-17

Family

ID=7876022

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1999/002327 WO2000008195A1 (fr) 1998-07-31 1999-07-30 Procede et dispositif de transfert d'oligonucleotides dans des cellules

Country Status (3)

Country Link
AU (1) AU6324199A (fr)
DE (1) DE19834612A1 (fr)
WO (1) WO2000008195A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001048181A3 (fr) * 1999-12-23 2002-04-18 Dornier Medizintechnik Procede pour transferer des molecules dans des cellules

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10223196B4 (de) * 2002-05-24 2004-05-13 Dornier Medtech Systems Gmbh Verfahren und Einrichtung zum Transferieren von Molekülen in Zellen
WO2020085281A1 (fr) * 2018-10-26 2020-04-30 国立大学法人九州大学 Procédé de projection de bulles, dispositif de projection de bulles et appareil de projection de bulles

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0137504A2 (fr) * 1983-10-13 1985-04-17 Rikagaku Kenkyusho Méthode et appareil pour implanter une substance étrangère dans des cellules vivantes
US4750100A (en) * 1986-06-06 1988-06-07 Bio-Rad Laboratories Transfection high voltage controller
WO1989002464A1 (fr) * 1987-09-07 1989-03-23 Amersham International Plc Modification de cellules vivantes
WO1991000358A1 (fr) * 1989-06-29 1991-01-10 Danisco A/S Procede d'introduction de molecules, notamment de matiere genetique, dans des cellules vegetales
US5098843A (en) * 1987-06-04 1992-03-24 Calvin Noel M Apparatus for the high efficiency transformation of living cells
EP0506632A2 (fr) * 1991-03-28 1992-09-30 Ente per le nuove tecnologie, l'energia e l'ambiente ( ENEA) Microporation par laser
WO1994009145A1 (fr) * 1992-10-13 1994-04-28 Cangene Corporation Transfection de particucles: procede de transfert de molecules polynucleotidiques dans des cellules
WO1997040679A1 (fr) * 1996-05-01 1997-11-06 Imarx Pharmaceutical Corp. Procedes d'apport de composes dans une cellule
US5753477A (en) * 1996-03-19 1998-05-19 University Technology Corporation Magneto-biolistic methods

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IL92529A0 (en) * 1989-12-03 1990-08-31 Yissum Res Dev Co Generation of transgenic vertebrates by employing transformed sperm cells via artificial insemination
US5766901A (en) * 1995-05-04 1998-06-16 The Board Of Trustees Of The Leland Stanford Junior University Apparatus and method for delivering a nucleotide into cell nuclei

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0137504A2 (fr) * 1983-10-13 1985-04-17 Rikagaku Kenkyusho Méthode et appareil pour implanter une substance étrangère dans des cellules vivantes
US4750100A (en) * 1986-06-06 1988-06-07 Bio-Rad Laboratories Transfection high voltage controller
US5098843A (en) * 1987-06-04 1992-03-24 Calvin Noel M Apparatus for the high efficiency transformation of living cells
WO1989002464A1 (fr) * 1987-09-07 1989-03-23 Amersham International Plc Modification de cellules vivantes
WO1991000358A1 (fr) * 1989-06-29 1991-01-10 Danisco A/S Procede d'introduction de molecules, notamment de matiere genetique, dans des cellules vegetales
EP0506632A2 (fr) * 1991-03-28 1992-09-30 Ente per le nuove tecnologie, l'energia e l'ambiente ( ENEA) Microporation par laser
WO1994009145A1 (fr) * 1992-10-13 1994-04-28 Cangene Corporation Transfection de particucles: procede de transfert de molecules polynucleotidiques dans des cellules
US5753477A (en) * 1996-03-19 1998-05-19 University Technology Corporation Magneto-biolistic methods
WO1997040679A1 (fr) * 1996-05-01 1997-11-06 Imarx Pharmaceutical Corp. Procedes d'apport de composes dans une cellule

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
BAO ET AL.: "In Vivo Transfection of Melanoma Cells by Lithotripter Shock Waves", CANCER RES., vol. 58, 15 January 1998 (1998-01-15), pages 219 - 221, XP002125162 *
DELIUS ET AL.: "Extracorporeal shock waves for gene therapy?", LANCET, vol. 345, 27 May 1995 (1995-05-27), pages 1377, XP002125161 *
LAUER ET AL.: "Shock wave permeabilization as a new gene transfer system in vitro.", J. CELL. BIOCHEM. SUPPL., vol. 0, no. 21A, 1995, pages 396, XP002125160 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001048181A3 (fr) * 1999-12-23 2002-04-18 Dornier Medizintechnik Procede pour transferer des molecules dans des cellules

Also Published As

Publication number Publication date
AU6324199A (en) 2000-02-28
DE19834612A1 (de) 2000-02-24

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