Method of constructing nucleus-implanted egg,...

Multicellular living organisms and unmodified parts thereof and – Method of making a transgenic nonhuman animal – Via microinjection of a nucleus into an embryo – egg cell – or...

Reexamination Certificate

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C800S014000, C800S025000

Reexamination Certificate

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07659443

ABSTRACT:
Disclosed is a method for constructing a nucleus-implanted egg, a parthenogenetic embryo and for producing a parthenogenetic mammal each having 2 haploid genome sets originating in mammalian ova, and provides methods of constructing a nucleus-implanted egg having a haploid genome set derived from primitive ovarian follicle egg (ng ovum) and a haploid genome set from MII phase (second meiosis metaphase) egg (fg ovum), a parthenogenetic embryo and a parthenogenetic mammal, including steps (1) introducing ng ovum into a nucleus-deleted deleted germinal vesicle stage (GV) egg, developing the obtained egg to MII phase by in vitro maturing and culturing to prepare a first nucleus-implanted egg, and (2) extracting MII phase chromosome from the first nucleus-implanted egg and introducing it into other fg ovum to prepare a second nucleus-implanted egg, wherein a ng or fg ovum from which an imprinted gene undergoing epigenetic modification during sperm generation is used.

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Wu et al., Regulated expression of two sets of paternally imprinted genes is necessary for mouse parthenogenetic development to term, Reproduction. vol. 131, No. 3, 2006, pp. 481-488. XP 002501736, ISSN: 1470-1626.
Kawahara et al., Nature Biotechnology, vol. 25, No. 9, Sep. 2007, pp. 1045-1050. XP-002501737.
Kawahara et al., Nature Protocols, Nature Publishing Group, UK, vol. 3 , No. 2, 2008, pp. 197-209. XP001538923.
T. Kono, Cytogenetic and Genome Research, vol. 113, No. 1-4, 2006, pp. 31-35, XP009107587.

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