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Definition: Transgenic Animal

Definition: Transgenic Animal. Animal in which a segment of DNA has been physically inserted into the genome. The genome of all cells of the organism contains the DNA segment. The DNA segment is present in the germ line so it can be transmitted to progeny as a Mendelian trait.

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Definition: Transgenic Animal

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  1. Definition: Transgenic Animal • Animal in which a segment of DNA has been physically inserted into the genome. • The genome of all cells of the organism contains the DNA segment. • The DNA segment is present in the germ line so it can be transmitted to progeny as a Mendelian trait.

  2. Retroviral Mediated Gene Transfer Mouse embryos MoMuLV Producing Cells Insertion of viral DNA containing transgene into embryos Transgenic Mouse

  3. PMS + HCG to superovulate 1-cell pronuclear stage mouse embryo. X 2-cell stage embryo Injection Pipette Holding Pipette Oviduct Transfer + +  Pregnant Foster Mother

  4. F e r t i l i z a t i o n R e i m p l a n t a t i o n M a t i n g M i c r o i n j e c t i o n C e l l D i v i s i o n G e s t a t i o n Events During Transgenesis E 1 E 2 E 2 1 D A Y 1 2 3 4 5 B i r t h E m b r y o R e c o v e r y PMS hCG 1 2 A M 1 2 A M 1 2 A M 1 2 A M 1 2 A M 1 2 A M 1 2 A M

  5. Embryonic Stem Cells • Pluripotent stem cells isolated form the inner cell mass of a blastocyst stage mouse embryo. • Cells retain ability to differentiate into all organs and cells. • Cells can be propagated under conditions in vitro that allow them to retain their totipotency- prevent differentiation in vitro. • Can be induced to differentiate in vitro into a number of cell types. ES cells can mimic early mammalian development in vitro. • Can be manipulated in vitro to alter the genome through homologous recombination.

  6. Pure Population of Targeted ES Cells ES cell Culture Targeted ES Cells Targeted ES Cells Injected into Blastocyst Uterine Transfer Chimera  Pregnant Foster Mother Blastocyst derived from mouse with white coat. X Germline transmission of ES cell-derived genome. Breed to WT white mouse

  7. B C D E F G E F G B C D Gene Knockout by Homologous RecombinationUsing an “O” or “Insertion” Type Construct A B C D E F G H I X neo A B C D E F G H I neo

  8. B C F G C F Gene Knockout by Homologous RecombinationUsing an “W” or “Replacement” Type Construct A B C D E F G H I X X TK neo A B G H I neo

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