Apparatus for genetic transformation

• US 5,149,655 A
• Filed: 06/21/1990
• Issued: 09/22/1992
• Est. Priority Date: 06/21/1990
• Status: Expired due to Term

• Current Assignee: PowderJect Vaccines, Inc. (Bayer AG)
• Original Assignee: Agracetus Incorporated
• Inventors: McCabe, Dennis E., Martinell, Brian J.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Beisner, William H.
• Application Number:
  US07/541,563
• Time in Patent Office: 824 Days
• Field of Search: 435/287, 435/173, 435/172.1-172.3, 435/313, 935/52, 935/53, 935/85, 604/68-70, 604/72, 604/140, 604/141, 604/146, 128/24 EL, 73/12, 73/167, 239/81
• US Class Current: 604/57
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

An apparatus for the particle mediated genetic transformation of organisms in vivo has two parts, a support unit and a hand unit. The hand unit is of a convenient hand manipulable size so that it can be placed readily against the organism the cells of which are to be transformed. The hand unit is connected to the support unit by a flexible umbilical so that the hand unit can be placed where desired. The apparatus is particularly well adapted for the convenient transformation of somatic cells of whole animals or humans.

Claims

1. An apparatus for performing genetic transformation of a living organism in vivo by accelerating carrier particles coated onto a carrier sheet into the organism comprising

a manually manipulable handle;

a body member mounted on the handle and of a size and shape so as to be readily carried and manually handled;

a carrier sheet;

the body member having an interior and a spark discharge chamber formed in the interior, the spark discharge chamber including a carrier sheet mounting location for mounting the carrier sheet thereon, the spark discharge chamber constructed so to present a shock wave to the carrier sheet mounting location;

a pair of electrodes extending into the body member and into the discharge chamber;

a carrier sheet retaining means removably attachable to the body member and located over the carrier sheet mounting location;

said carrier sheet retaining means including

a retaining screen located spaced from the carrier sheet so that the carrier sheet will impact the retaining screen when accelerated from the carrier sheet mounting location;

a support unit including a source of electrical discharge energy; and

an umbilical connecting the electrode to the support unit, so that electrical energy can be supplied to the electrodes for initiating an electric spark discharge between the electrodes while permitting the body member to be readily placed adjacent the organism to be transformed.

2. An apparatus as claimed in claim 1 wherein the body member has a front and the spark discharge chamber opens onto the front of the body member and wherein carrier sheet mounting location is a recessed step located surrounding an opening of the discharge chamber onto the front of the body member.

3. An apparatus as claimed in claim 1 wherein the carrier sheet retaining means includes a slot through which the retaining screen is inserted to be mounted in the carrier sheet retaining means.

4. An apparatus as claimed in claim 1 wherein the support unit includes a source of helium, wherein the carrier sheet retaining means includes an interior and a gas port through which gas may flow into the interior, and wherein the umbilical further includes means to conduct helium from the support unit to the gas port on the carrier sheet retaining means.

5. An apparatus as claimed in claim 1 wherein the body member includes ann opening into the discharge chamber adjacent to the electrodes and wherein there is further an end plate removably attached to the body member to cover the opening adjacent the electrodes so that convenient access can be had to the electrodes.

6. An apparatus for performing a genetic transformation of a living organism by the acceleration of carrier particles comprising:

a support unit constructed and arranged so as to supply a motive force for accelerating carrier particles into the organism; and

hand-held means comprising a body member, a carrier sheet, and a retaining screen for retaining the carrier sheet attached to the body member, the body member having formed in it a discharge chamber constructed so as to generate a shock wave from said motive force and to direct shock wave therein toward the carrier sheet and constructed so as to hold the carrier sheet thereon in such a way that the shock wave will be applied to accelerate the carrier sheet, the retaining screen arranged so as to retain the carrier sheet after it is accelerated so that the carrier particles are separated therefrom; and

umbilical means for transmitting the motive force from the support unit to the hand-held unit.

7. An apparatus as claimed in claim 6 wherein the motive force supplied by the support unit is electrical potential, and wherein the body member includes means therein to convert the electrical potential to physical force to accelerate the carrier sheet.

8. An apparatus as claimed in claim 7 wherein the means to convert the electrical force to physical force includes a pair of spaced electrodes which are bridged by a spark discharge when sufficient electrical potential is applied between them.

9. An apparatus as claimed in claim 6 wherein the support unit further includes a source of helium, wherein the umbilical means further includes means to conduct the helium to the hand-held unit, and wherein the hand-held unit further includes means to conduct the helium to the path of travel of the carrier sheet.

10. An apparatus as claimed in claim 6 wherein the retaining screen is removably attached to the body member so that the carrier sheet may be placed in the body member.