METHOD FOR ACQUIRING GENETICALLY IDENTICAL GAMETE FROM LETHAL FISH HAPLOID-DERIVED GERM CELL VIA GERM LINE CHIMERA

• US 20120304322A1
• Filed: 02/09/2011
• Published: 11/29/2012
• Est. Priority Date: 02/09/2010
• Status: Active Grant

• Current Assignee: National University Corporation Hokkaido University
• Original Assignee: National University Corporation Hokkaido University
• Inventors: Yamaha, Etsuro, Arai, Katsutoshi, Fujimoto, Takafumi, Saito, Taiju
• Granted Patent: US 8,921,643 B2
• Time in Patent Office: Days
• US Class Current: 800/20
• CPC Class Codes: A01K 2207/05 Animals modified by non-integrating nucleic acids, e.g. antisense, RNAi, morpholino, episomal vector, for non-therapeutic purpose
• CPC Class Codes: A01K 2227/40 Fish
• CPC Class Codes: A01K 67/027 New or modified breeds of vertebrates
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: C12N 15/09 Recombinant DNA-technology
• CPC Class Codes: C12N 15/873 Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
• CPC Class Codes: C12N 5/10 Cells modified by introduction of foreign genetic material

Abstract

Disclosed are a method for acquiring a germ line chimeric fish having fish haploid germ cells, a germ line chimeric fish having haploid germ cells obtained by the aforesaid method, and a genetically identical gamete, said gamete having been derived from a donor haploid germ cell, produced by a germ line chimeric fish obtained by the aforesaid method.

Claims

1. A method for acquiring a chimeric fish having a haploid germ cell, said fish being selected from the group consisting of loach, goldfish and zebrafish, comprising the steps of:

a) genetically inactivating a fish egg or a fish sperm, followed by insemination to provide a haploid donor fertilized egg;

b) obtaining a fertilized egg of a host fish; and

c) transplanting a donor primordial germ cell obtained from a haploid donor embryo developed from the haploid donor fertilized egg into a host embryo derived from the host fertilized egg.

5. The method according to claim 1, wherein the genetic inactivation of an egg or a sperm is performed by ultraviolet irradiation or radiation irradiation.

6. The method according to claim 1, wherein the host fish is a sterile host fish obtained by the fertilization of a normal polyploid egg by a diploid sperm of a tetraploid fish.

7. The method according to claim 1, wherein the donor is a color mutant or a genetic recombinant.

8. The method according to claim 1, wherein the method for sterilizing the host fish involves knocking down a dead end gene in the host fertilized egg.

9. The method according to claim 8, wherein the knocking down is injecting a dead end antisense morpholino oligonucleotide into the host fertilized egg.

13. The method according to claim 1, wherein the transplantation is carried out by a blastomere transplantation method, a single primordial germ cell transplantation method, a poly primordial germ cell transplantation method, or a blastoderm transplantation method.

14. The method according to claim 1, wherein the donor or host fertilized egg is obtained, followed by removing the membrane of the egg.

15. A method for acquiring genetically identical gametes by the method according to claim 1.

16. Genetically identical gametes obtained by the method according to claim 15.

2-4. -4. (canceled)

10-12. -12. (canceled)

17. A chimeric host fish having a haploid primordial germ cell derived from a haploid germ cell.

18. The chimeric host fish according to claim 17, wherein the chimeric host fish is a sterilized individual.

19. Genetically identical gametes obtained from the chimeric host fish according to claim 17.

20. A haploid clonal fish gamete derived from a haploid primordial germ cell.

21. (canceled)

22. A chimeric fish embryo from a host fish embryo transplanted with a haploid donor-derived haploid primordial germ cell.

23. (canceled)