Abstract: Disclosed is a novel means which makes it possible to steadily mass-produce knockout individuals even in large animals. The method of the present invention is a method for producing a non-human large mammal or fish (non-human animal) that produces gametes originating in a different individual, and comprises transplanting at least one pluripotent cell derived from a second non-human animal into an embryo derived from a first non-human animal, said embryo being at a cleavage stage and having a genome in which a function of nanos3 gene is inhibited, to prepare a chimeric embryo, and allowing said chimeric embryo to develop into an individual.

Abstract: Disclosed is a novel means which enables satisfactory preservation of embryos and fertilized eggs in the non-frozen state for a longer period than conventional means, which novel means also achieves high hatching ability and a high conception rate of the embryos after the preservation. The method for preserving a mammalian embryo(s) or fertilized egg(s) of the present invention comprises immersing a mammalian embryo(s) or fertilized egg(s) in a medium containing 20 to 80% (v/v) serum and 10 to 100 mM Good's buffer, and storing the embryo(s) or fertilized egg(s) at non-freezing low temperature. The preservative solution for a mammalian embryo(s) or fertilized egg(s) of the present invention essentially consists of a medium containing 20 to 80% (v/v) serum and 10 to 100 mM Good's buffer. The Good's buffer is preferably HEPES.

Abstract: Disclosed is a novel means which makes it possible to steadily mass-produce knockout individuals even in large animals. The method of the present invention is a method for producing a non-human large mammal or fish (non-human animal) that produces gametes originating in a different individual, and comprises transplanting at least one pluripotent cell derived from a second non-human animal into an embryo derived from a first non-human animal, said embryo being at a cleavage stage and having a genome in which a function of nanos3 gene is inhibited, to prepare a chimeric embryo, and allowing said chimeric embryo to develop into an individual. When a pluripotent cell having a genome in which a desired gene is knocked out is used as the pluripotent cell derived from the second non-human animal, the first non-human animal capable of producing germ cells in which the desired gene is knocked out is obtained, and therefore knockout non-human animals can easily be mass-produced by mating such non-human animals.

Abstract: Disclosed is a novel means which enables satisfactory preservation of embryos and fertilized eggs in the non-frozen state for a longer period than conventional means, which novel means also achieves high hatching ability and a high conception rate of the embryos after the preservation. The method for preserving a mammalian embryo(s) or fertilized egg(s) of the present invention comprises immersing a mammalian embryo(s) or fertilized egg(s) in a medium containing 20 to 80% (v/v) serum and 10 to 100 mM Good's buffer, and storing the embryo(s) or fertilized egg(s) at non-freezing low temperature. The preservative solution for a mammalian embryo(s) or fertilized egg(s) of the present invention essentially consists of a medium containing 20 to 80% (v/v) serum and 10 to 100 mM Good's buffer. The Good's buffer is preferably HEPES.

Abstract: The novel means by which an efficiency of ovum collection can be easily determined in bovine at gene level is disclosed. The present inventors performed the genomic linkage analysis using bovine populations with high and low efficiency of ovum collection and to identify GRIA1 gene, which encodes an ion channel protein, as a factor deeply related to an efficiency of ovum collection. Bovines having a mutation (e.g. the amino acid substitution of aa306) in GRIA1 produce significantly fewer ova on superovulatory treatment than those not having the mutation. Therefore, the efficiency of ovum collection can be determined based on the existence of a mutation in GRIA1 gene.

Abstract: The novel means by which an efficiency of ovum collection can be easily determined in bovine at gene level is disclosed. The present inventors performed the genomic linkage analysis using bovine populations with high and low efficiency of ovum collection and to identify GRIA1 gene, which encodes an ion channel protein, as a factor deeply related to an efficiency of ovum collection. Bovines having a mutation (e.g. the amino acid substitution of aa306) in GRIA1 produce significantly fewer ova on superovulatory treatment than those not having the mutation. Therefore, the efficiency of ovum collection can be determined based on the existence of a mutation in GRIA1 gene.