Method for developing transgenic mice

• US 6,492,575 B1
• Filed: 12/28/1999
• Issued: 12/10/2002
• Est. Priority Date: 08/13/1996
• Status: Expired due to Term

• Current Assignee: Boehringer Ingelheim International GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Original Assignee: Boehringer Ingelheim International GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Inventors: Wang, Zhao-Qi, Wagner, Erwin
• Primary Examiner(s): Priebe, Scott D.
• Assistant Examiner(s): Paras, Jr., Peter
• Application Number:
  US09/242,406
• Publication Number:
  US 20020178457A1
• Time in Patent Office: 1,078 Days
• Field of Search: 435/288.3, 435/305.1, 435/455, 800/14, 800/7, 800/25, 800/18, 800/8, 800/13, 800/21
• US Class Current: 800/25
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic

Abstract

Process for producing mammals with defined genetic properties, particularly transgenic mammals. Animals which are derived completely from ES cells are obtained by injecting totipotent cells (embryonic stem cells or embryonic germ cells) genetically manipulated and cultured in vitro into tetraploid blastocysts and implanting the resulting embryo in a foster mother, in a single operation.

Claims

1. A method for producing a transgenic mouse with a desired genotype, said method comprising:

(a) obtaining a mouse tetraploid blastocyst;

(b) introducing a plurality of mouse totipotent cells into said tetraploid blastocyst, wherein said totipotent cells have been genetically manipulated to comprise a transgene conferring a desired genotype to said cells, thereby producing a transgenic embryo; and

(c) transferring said embryo into a foster mother under conditions favoring the development of said embryo into a transgenic mouse comprising said desired genotype.

2. The method of claim 1, wherein said blastocyst is obtained by electrofusion of two-cell embryos and subsequent culturing.

3. The method of claim 1, wherein said totipotent cells are embryonic stem cells.

4. The method of claim 1, wherein said totipotent cells are embryonic germ cells.

5. The method of claim 1, wherein said totipotent cells are introduced into said tetraploid blastocyst by microinjection.

6. The method of claim 1, wherein said totipotent cells were obtained by the introduction of plasmids which carry the desired genetic modification.

7. The method of claim 1, wherein said totipotent cells comprise at least one transgene.

8. A method for producing a transgenic mouse, comprising:

(a) obtaining a mouse tetraploid blastocyst by electrofusion of two-cell diploid embryos obtained from mouse;

(b) genetically manipulating mouse embryonic stem cells, wherein said genetic manipulation introduces a transgene into said embryonic stem cells; and

(c) introducing one or more of said embryonic stem cells into said tetraploid blastocyst, thereby generating a transgenic embryo; and

(d) implanting said transgenic embryo into a foster mother under conditions favoring development of said embryo into a transgenic mouse.

9. The method of claim 8, wherein said embryonic stem cells are R1 cells.

10. The method of claim 8, wherein said embryonic stem cells are GS1 cells.

11. The method of claim 8, wherein said embryonic stem cells are J1 cells.

12. The method of claim 8, wherein said embryonic stem cells are TT2 cells.

13. The method of claim 8, wherein said embryonic stem cells are introduced into said tetraploid blastocyst by microinjection.

14. The method of claim 8, wherein said embryonic stem cells comprise at least one transgene.