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Production of ungulates, preferably bovines that produce human immunoglobulins

  • US 20050183145A1
  • Filed: 12/14/2004
  • Published: 08/18/2005
  • Est. Priority Date: 11/19/1999
  • Status: Abandoned Application
First Claim
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1. A method for producing a cloned ungulate wherein the expression of both copies of a gene essential for B cell production has been knocked out, selected from the group consisting of Igα

  • , IgM, E2A, EBF, BSAP, rag-1 and rag-2, which comprises the following steps;

    (i) producing a male and/or female ungulate cell wherein the expression of one or both copies of the Igα

    , E2A, EBF, BSAP, IgM heavy chain, rag-1 and/or rag-2 gene has been eliminated by targeted disruption;

    (ii) using said cell or DNA therefrom as a donor for nuclear transfer by fusing or inserting said donor cell or nucleus into an oocyte or blastomere, which is enucleated before or after transfer, activating the resulting nuclear transfer unit and/or the oocyte prior or simultaneous to nuclear transfer and culturing in a suitable medium to produce a nuclear transfer embryo;

    (iii) introducing said nuclear transfer embryo into a female ungulate; and

    (iv) obtaining a cloned fetus or animal ungulate that expresses the genotype of the donor differentiated cell, in which one or both copies of the IgM (mu) chain, Igα

    , E2A, EBF, BSAP, rag-1 and/or rag-2 gene have been eliminated; and

    (v) optionally, mating said cloned male or female ungulate with another cloned female ungulate wherein one copy of the IgM, rag-1 or rag-2 gene has been knocked out and selecting progeny wherein both copies of the Igα

    , E2A, EBF, BSAP, IgM, rag-1 or rag-2 genes have been knocked out.

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