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Alison Van Eenennaam, Ph.D. Cooperative Extension Specialist Animal Biotechnology and Genomics Department of Animal Science University of California, Davis, USA

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Alison Van Eenennaam, Ph.D. Cooperative Extension Specialist Animal Biotechnology and Genomics Department of Animal Science University of California, Davis, USA Uso de la edición genética para incorporar alelos iatrogénicos o intergénicos deseables que aumenten la ganancia genética en los programas de mejoramiento de animales de carne Alison Van Eenennaam, Ph.D. Cooperative Extension Specialist Animal Biotechnology and Genomics Department of Animal Science University of California, Davis, USA Email: [email protected] Twitter: @BioBeef Photo credit: Cornell Alliance for Science http://animalscience.ucdavis.edu/animalbiotech Uruguay 10/10/2016 Animal Genomics and Biotechnology Education La FDA no regula la mejora animal por cruzamiento o por tecnologías de reproducción asistida (Solo el uso de drogas ) Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Que es la edición genómica? Meganuclease Site- Zinc finger directed TALENs nuclease CRISPR/Cas9 Double Non-homologous stranded break Homologous end joining (NHEJ) recombination http://genok.no/wp-content/uploads/2015/06/250615_Emerging_technologies_final.pdf Uruguay 10/10/2016 Animal Genomics and Biotechnology Education La mejora por mutación en plantas introduce cortes en el ADN. Esta forma de mejoramiento ha sido utilizada desde 1950 y a escapado a la legislación hasta la fecha. Se han liberado mas de 3200 cultivos mejorados mediante esta tecnología, entre ellos mas de 600 variedades de trigo, arroz y maiz Uruguay 10/10/2016 Photo by Stephen Ausmus, USDA Animal Genomics and Biotechnology Education Selección por cruzamiento de ganado con doble musculo (mutación natural del gen de la miostatina) Uruguay 10/10/2016 Edición genomica del gen de la muostatina para obtener doble musculo en ganado (sustitución de alelos interespecifica) Proudfoot C, et al. 2015. Genome edited sheep and cattle. Transgenic Res. 2015 Feb;24(1):147-53. Uruguay 10/10/2016 Mejoramiento genético (permanente, cumulativo) en remplazo al uso de antibióticos/químicos para el manejo de enfermedades Uruguay 10/10/2016 Animal Genomics and Biotechnology Education • African Swine Fever • Porcine Reproductive and Respiratory Syndrome (PRRSV) virus Lillico et al. 2016. Mammalian interspecies substitution of immune modulatory alleles by genome editing. Sci Rep 6:21645. Whitworth et al. 2016. Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus (PRRSV). Nature Biotechnology 34:20-22. Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Los animales resistentes a enfermedad repercuten en el bienestar animal y la salud humana Van Eenennaam, A.L. and A.E. Young. 2015. Animal agriculture and the importance of agnostic governance of biotechnology. Agriculture and Food Security 4:21 Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Terneros sin cuernos genéticamente editados Naturally-occurring bovine allele at polled locus Carlson DF, Lancto CA, Zang B, Kim E-S, Walton M, et al. 2016. Production of hornless dairy cattle from genome-edited cell lines. Nat Biotech 34: 479-81 Uruguay 10/10/2016 Animal Genomics and Biotechnology Education Características de interés para ser modificadas en ganado de carne. • Polled/Horned • Myostatin • Recessive Genetic Conditions • Tenderness loci (calpastatin/calpain) • GH gene • SCD (stearoyl-CoA desaturase) Editing would synergistically accelerate conventional breeding programs (not replace them!) by precisely bringing in discrete desired genetic variation as needed Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Tasa acelerada de ganancia genética en características cuantitativas cuando se realizan de 1-20 modificaciones genéticas Jenko, J. et a. 2015. Potential of promotion of alleles by genome editing to improve quantitative traits in livestock breeding programs. Genetics Selection Evolution 47: 1-14. Uruguay 10/10/2016 Animal Genomics and Biotechnology Education Kasinathan, P. et al. 2015. Acceleration of genetic gain in cattle by reduction of generation interval. Sci. Rep. 5, 8674; DOI:10.1038/srep08674 Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Como se puede integrar la modificacion genetica en los programas de mejoramiento? Van Eenennaam, A. L. 2017. Genetic Modification of Food Animals. Current Opinion in Biotechnology. Submitted. Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Llegara la edición genética a disparar el control regulatorio? Si/No/Tal vez The Cartagena Protocol defines “Living modified organism” to mean any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology "Modern biotechnology" means the application of: a. In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA) and direct injection of nucleic acid into cells or organelles, or b. Fusion of cells beyond the taxonomic family, that overcome natural physiological reproductive or recombination barriers and that are not techniques used in traditional breeding and selection Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Gene editing of myostatin to obtain double muscle Nelore cattle – same allele as occurs “naturally” - What is the “novel combination of genetic material”? Proudfoot C, et al. 2015. Genome edited sheep and cattle. Transgenic Res. 2015 Feb;24(1):147-53. Uruguay 10/10/2016 Animal Biotechnology and Genomics Education La edicion genomica puede introducir cambios en el ADN que pueden introducir transgenes o no Meganuclease Zinc finger TALENs CRISPR/Cas9 Sander JD, Joung JK. CRISPR-Cas systems for editing, regulating and targeting genomes. Nat Biotech 2014;32:347-355. Uruguay 10/10/2016 Animal Genomics and Biotechnology Education La FDA regula los animales geneticamente editados en EEUU The Food and Drug Administration’s Center for Veterinary Medicine (CVM) evaluates GE animals under the new animal drug provisions of the Federal Food Drug and Cosmetic Act (FFDCA). The act defines drugs as “articles (other than food) intended to affect the structure or any function of the body of man or other animals.” The rDNA construct in the resulting GE animal is thus a regulated article that meets the drug definition; the GE animal itself is not a drug. The FDA defines “genetically engineered (GE) animals” as those animals modified by rDNA techniques, including the entire lineage of animals that contain the modification, and regulates based on the use of rDNA techniques. All GE animals are captured under these provisions, regardless of their intended use. Thus although the review is product based, the process used to produce the genetic change that results in the product (e.g. rDNA versus traditional breeding) has implications for triggering regulatory oversight. Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Son novedosos los terneros sin cuernos geneticamente editados ? Naturally-occurring bovine allele at polled locus What is the “new animal drug” in this case? Tan et al. 2013. Efficient nonmeiotic allele introgression in livestock using custom endonucleases. PNAS 110: 16526-31. Uruguay 10/10/2016 Animal Biotechnology and Genomics Education El uso de la edicion genetica vs. cruza selectiva para introducir el alelo natural “sin-cuernos” en el ganado Holando. Attribute Polled Holstein Polled Holstein through gene through editing introgression Phenotype: No horns YES YES Mutation uniquely detectable NO NO Food safety concerns NO NO associated with phenotype # generations taken to achieve ONE MANY polled >15/16 Holstein (FAST) (SLOW) Linkage drag? NO YES Improved animal welfare YES YES Regulated? Depends on definition of NO regulated article Likely to happen Not if takes years and NO costs millions of dollars Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Tener una regulación diferente para terneros sin cuernos lecheros o de carne, tiene sentido? Carroll D, Van Eenennaam AL, Taylor JF, Seger J, Voytas DF. 2016. Regulate genome- edited products, not genome editing itself. Nat Biotech 34: 477-9 rdcu.be/hUVn Uruguay 10/10/2016 Animal Biotechnology and Genomics Education Que ocurre en la Union Europea ? GMO (included in Directive 2001/18/EC) is defined as an „organism, with the exception of human beings, in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination‟. The Annexes of the Directive says techniques of genetic modification includes: i)recombinant nucleic acid techniques involving the formation of new combinations of genetic material, ii) direct introduction into an organism of heritable material prepared outside the organism iii) cell fusion Techniques of genetic modification yielding organisms to be excluded from the Directive include, for example, those derived by mutagenesis. Techniques not considered to result in a genetic modification include in vitro fertilisation, “natural” processes like conjugation, transduction, transformation and polyploidy induction. Uruguay 10/10/2016 Animal Biotechnology and Genomics Education If not an organism Organism 1. If Human Non-Human Organism 2. PART 1. If not altered Its Genetic Material is Altered Product (>20 bp foreign genetic material) 3. (the result) Is capable of occurring naturally by mating &/or natural recombination The Genetic Alteration 4. Is NOT capable of occurring naturally by mating &/or natural selection Uruguay 10/10/2016 Animal BiotechnologyAnimal Genomics and Genomics and Biotechnology Education Education Por que 20 bp? Based on statistical calculations, the Joint Research Centre (2011) has arrived at a threshold for new combinations of genetic material of 20 nucleotides for plant genomes.
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