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PRIMARY CELL-MEDIATED GENE TRANSFER - Gene transfer by embryo cloning -

AC21 Advisory Committee Meeting November 28, 2007. PRIMARY CELL-MEDIATED GENE TRANSFER - Gene transfer by embryo cloning -. Jim Robl Hematech, Inc. DNA TRANSFER BY PRIMARY CELL CLONING. Primary cell – Cell with a finite lifespan. Transfection. Selection. Primary fibroblast cells.

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PRIMARY CELL-MEDIATED GENE TRANSFER - Gene transfer by embryo cloning -

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  1. AC21 Advisory Committee Meeting November 28, 2007 PRIMARY CELL-MEDIATED GENE TRANSFER- Gene transfer by embryo cloning - Jim Robl Hematech, Inc.

  2. DNA TRANSFER BY PRIMARY CELL CLONING Primary cell – Cell with a finite lifespan Transfection Selection Primary fibroblast cells Transgenic cells Vector Embryonic cloning Enucleated oocyte

  3. MULTIPLE GENETIC MODIFICATIONS Transfection and selection Primary Clone to Rejuvenate Cell line establishment Clone to make Calves Transfection and selection 1st Modification Clone to Rejuvenate Cell line establishment Clone to make Calves Transfection and selection 2nd Modification Clone to Rejuvenate Additional modifications

  4. GENE TARGETING STOP SEQUENCE (GENE KNOCKOUT) SELECTION SEQUENCE Homologous arms DNA Sequence (chromosome) Gene Sequence Recombination 1st Chromosome 2nd Chromosome

  5. GENE KNOCKOUT CATTLE PRION GENE KNOCKOUT CATTLE IMMUNOGLOBULIN GENE KNOCKOUT CATTLE

  6. MICROCHROMOSOME TRANSFER Chromosome fragment Chromosome GENE SEQUENCE OF INTEREST REMOVE UNWANTED GENES Transgenic cells

  7. TRANSCHROMOSOMIC CALVES

  8. THERAPEUTIC PROTEINS PRODUCED IN MILK Niemann, H. & W.A. Kues. Reprod Fertil Dev 2007; 19: 762-770.

  9. PIG-TO-HUMAN ORGAN TRANSPLANTATION Niemann, H. & W.A. Kues. Reprod Fertil Dev 2007; 19: 762-770.

  10. TRANSGENIC LIVESTOCK FOR AGRICULTURE Niemann, H. & W.A. Kues. Reprod Fertil Dev 2007; 19: 762-770.

  11. SUMMARY • Technologies exist to make complex genetic modifications in livestock species • Commercial effort focused on human therapeutic applications • One product currently approved for marketing • Agricultural applications at research level

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