Mammalian artificial chromosomes and methods of using same
AssigneeThe Regents of the University of California
InventorImmo E. Scheffler
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Abstract
The present invention provides a mammalian artificial chromosome (MAC), comprising a centromere and a unique cloning site, said MAC containing less than 0.1% of the DNA present in a normal haploid genome of the mammalian cell from which the centromere was obtained. The invention further provides a MAC, wherein the unique cloning site is a nucleic acid sequence encoding a selectable marker. The invention also provides methods of preparing a MAC. In addition, the invention provides methods of stably expressing a selectable marker in a cell, comprising introducing a MAC containing the selectable marker into the cell. The invention also provides a cell containing a MAC expressing an exogenous nucleic acid sequence and a transgenic mammal expressing a selectable marker.
Claims14
A method of producing a mouse expressing a selectable marker which is present on a mammalian artificial chromosome (MAC), comprising the steps of:
- a. introducing a MAC containing the selectable marker into an ovum cell, said ovum cell fertilized into a zygote at the time of or following said introduction of said MAC, wherein said zygote contains said MAC containing said selectable marker;
- b. implanting said MAC-containing zygote into a female mammal;
- and
- c. selecting a mouse produced from said zygote, wherein said mouse expresses said selectable marker.
A method of producing a mouse expressing a selectable marker which is present on a mammalian artificial chromosome (MAC), comprising the steps of:
- a. introducing a MAC containing the selectable marker into an embryonic stem cell;
- b. introducing said MAC-containing embryonic stem cell into an embryo;
- c. implanting said MAC-containing embryo into a female mammal;
- and
- d. selecting a mouse produced from said embryo, wherein said mouse expresses said selectable marker.
The method of claim wherein said MAC is less than about 0.1% of the size of the normal haploid genome of the mammalian cell from which the centromere was obtained.
A method of producing a transgenic mouse expressing a selectable marker which is present on a mammalian artificial chromosome (MAC), comprising the steps of:
- a. introducing a MAC containing the selectable marker into an embryonic stem cell by microcell fusion of an embryonic stem cell and a second cell which contains the MAC to produce a hybrid cell expressing the selectable marker;
- b. introducing said MAC-containing hybrid cell into an embryo;
- c. implanting said MAC-containing embryo into a female mammal;
- d. selecting a mouse produced from said embryo, wherein germ cells of said mouse express said selectable marker;
- f. mating said mouse with another mouse to produce progeny;
- and
- g. selecting a transgenic mouse from said progeny.
Dependencies are inferred from the claim text.