1. Biotechnological methods in animal production Peter Chrenek chrenekp@vuzv.sk

2. Introduction Modern biotechnology has emerged in the last two-three decades and it utilizes cells and biomolecules for the application in medicine, agriculture and protection of the environment. In Agriculture - Transgenesis - Nuclear transfer (cloning) - Cell culture and Tissue Ingeneering - Recombinant DNA

3. Introduction Animal Biotechnology - Genetic – genetic markers - genomic - Biodiversity – genetic resources - Nutrition – nutrigenomic - Welfare - Reproduction – artificial insemination - embryotransfer - cryopreservation - transgenesis „GMO“

4. Animal Biotechnology in Reproduction Transgenesis 1) basic research (early detection of integration and expression, targeted integration,....) 2) production of pharmaceutic (recombinant) substances 3) increase milk, meat production (yield) and health of animals 4) in animal nutrition 5) xenotransplantations 6) production of cloned transgenic embryos (HYBRID EMBRYOS) – possiblility to isolate and use of embryonic stem cells (BIOMEDICAL PURPOSE)

5. Animal Biotechnology in Reproduction Transgenesis 1) Early detection of targeted gene integration and expression, targeted integration, EGFP,....)

6. Animal Biotechnology in Reproduction Transgenesis Rabbit milk obtained by use of electrical device for milking and is drained into a collection container. Female of weight 4-5 kg produces ¼ liter of milk daily. Rabbit milk has a high fat content. 2) Production of pharmaceutic (recombinant) substances

7. Animal Biotechnology in reproduction – Transgenesis 2) Production of pharmaceutic (recombinant) substances Zhao Sihai et al., 2009 recombinant proteinUseauthors h alpha1-antitrypsinemohysemaMassoud et al., 1990 h IL-2?Buhler et al., 1990 h tPAthrombosisReigo et al., 1993 h erythropoietinanemiaRodriguez et al., 1995 h IGF-1GH deficiencyBrem et al., 1994 h ESDMischemiaStormqvist et al., 1997 h GHGH deficiencyHammer et al., 1985 h alpha-glucuronidaseglycogenBijvoet et al., 1999 salmon calcitoninosteoporosisMcKee et al., 1998 eCG?Galet, 2001 h NGF-betaneuropathyCoulibaly et al., 1999 h PChPC defficiencyChrenek et al., 2002 hFVIIIhemophiliaChrenek et al., 2003 hFVIIIhemophiliaHiripi et al., 2003 bochymosincheese productionBrem et al., 1995

8. Animal Biotechnology in Reproduction – Transgenesis Transgenic rabbits

9. Production and use of GM animals 3) Increase milk and meat production (yield) and animal health Belgian blue cattle – natural mutation of gene for myostatin ttp:www.hemmy.net/images/animals/supercow01.jpg 'Mighty mice' Comparison of body size and anatomical conformation between normal mouse (left) and genetically modified mouse with absence of myostatin and overproduction of follistatin (right). Block out the gene for myostatin – double muscling of farm animals

10. Zdroj: http://technet.idnes.cz/gmo-zvirata-vepr-prasata-0xo- /veda.aspx?c=A150704_224211_veda_mlahttp://technet.idnes.cz/gmo-zvirata-vepr-prasata-0xo- /veda.aspx?c=A150704_224211_veda_mla

11. Animal Biotechnology in reproduction - Transgenesis 3) Increase milk and meat production (yield) and animal health GM salmon up: a transgenic salmon is much bigger than normal individual at the same age of 18 months. down: fish size will equal at 36 months.

12. Animal Biotechnology in Reproduction - Transgenesis 5) xenotransplantations Pigs could be a source of new organs. In pig cells was detected a porcine endogenous retrovirus (PERV), which is one from more reasons, why people are so skeptical about xentortansplantation. http://www.thenakedscientists.com/HTML/uploads/pics/jamil1

13. Animal Biotechnology in Reproduction – Nuclear transfer 6) production of transgenic embryonic stem cells Cloning Blastocyst Embryoblast Cloning (therapeutic a reproductive) Hybrid embryos - oocyte from cattle and human somatic transgenic cell - oocyte from rabbit and human somatic transgenic cell - oocyte from rabbit and human somatic transgenic cell Embryonic stem cells

14. Animal Biotechnology in reproduction – Cell culture embryonic stem cells Pluripotent – ability to differentiate into all types of cells, tissues and organs

15. Thank you for your attention