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Overview of the Ethical Issues of Germ Line Modification in Animals 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 Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Overview of the Ethical Issues of Germ Line Modification in Animals Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Breeding Method: Selective Breeding Genomic Selection Mutagenesis (ENU) Gene Knockout (ES) Genetic engineering Sterile insect technique Cloning Gene Editing Species Research Millions of Mice/Laboratory Rodents/Zebrafish Van Eenennaam 12/8/2015 Biomedical Products Pharma products Pigs ‐ Rabbit ‐ Ruconest Goat – ATryn, spider silk Xenotransplantation Cattle – polyclonal human antibodies Pets GloFish Micropigs Pest Control TseTse fly – sleeping sickness Mosquitoes – malaria resistance Moths – agricultural pest control Agriculture AquAdvantage Salmon – fast growth Polled Holsteins Disease resistance Improved product quality Decrease environmental footprint Single gender offspring Animal Genomics and Biotechnology Education Attitudes towards the use of animals in research • People’s attitudes toward experiments involving animals are dependent upon the beneficiary, purpose or necessity of the research (are there non-animal alternatives?) • People are more supportive of using smaller-brained animals such as mice and rats and less supportive of higher order animals • Public concerns about the use of animals in research are accentuated when genetically modified (GM) animals are used • Concern about increase in the number of animals used in research due to the (previously) inefficient and unpredictable nature of the genetic modification process • The creation of GM animals for research may challenge the Three R’s principles (replacement, reduction, refinement) Ormandy, E.H. and C.A. Schuppli. 2014. Public Attitudes toward Animal Research: A Review. Animals 391-408 Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Factors affecting people’s acceptance of animals in research: Invasiveness, genetic modification, regulation • Support for non-invasive research (in this paper on pigs for agricultural research) was high irrespective of regulation • Became less acceptable if invasive, but acceptance increased if animals did not suffer unnecessarily, that the benefits outweighed the harms, that the research was regulated, and that the use of animals was reduced, refined (minimizing pain and suffering), and replaced wherever possible • Low level of support for research using GM animals, and proposing regulation had little effect on this • In the case of acceptability of GM animals in research only 26% of people weighed costs and harms in making a decision as compared to 72% who used cost-harm for invasiveness decision Ormandy, E.H., C.A. Schuppli, and D.M. Weary. 2015. Public Attitudes towards the Use of Animals in Research: Effects of Invasiveness, Genetic Modification, and Regulation. Anthtozoos Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Factors affecting people’s acceptance of animals in research: Species, breeding method used to achieve genetic modification • Participants who were willing to accept biomedical research involving genetic modification on zebrafish were also willing to support the same research on mice • Participants expressed low levels of support for ENU mutagenesis on zebrafish in both unregulated and regulated research programs • Some participants expressed a preference for use of geneticallymodified (GM) animals over ENU mutagenesis, based on the belief that the former involves less pain and improves both the accuracy and efficiency of the animal models Ormandy, E.H., C.A. Schuppli, and D.M. Weary. 2012. Factors Affecting People’s Acceptance of the Use of Zebrafish and Mice in Research. ATLA 321-333. Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Method Used: Selective Breeding Genomic Selection Mutagenesis (ENU) Gene Knockout (ES) Genetic engineering Sterile insect technique Cloning Gene Editing Species Research Millions of Mice/Laboratory Rodents Van Eenennaam 12/8/2015 Biomedical Products Pharma Products Pigs ‐ Rabbit ‐ Ruconest Goat – ATryn, spider silk Xenotransplantation Cattle – polyclonal human antibodies Pets GloFish Micropigs Pest Control TseTse fly – sleeping sickness Mosquitoes – malaria resistance Moths – agricultural pest control Agriculture AquAdvantage Salmon – fast growth Polled Holsteins Disease resistance Improved product quality Decrease environmental footprint Single gender offspring Animal Genomics and Biotechnology Education GLOFISH Species: Tetras, Danios and Barbs Breeding method: Genetic Engineering Purpose: Pet Purpose Should we keep pets? Does the trait impact fish? Should we breed animals to be pets? Species Does breeding method hurt fish? Is this a food species? Do they pose an environmental hazard? Governance? Van Eenennaam 12/8/2015 Does breeding method pose new risks? How else is this trait achieved in pet fish? Breeding Method Animal Genomics and Biotechnology Education Method Used: Selective Breeding Genomic Selection Mutagenesis (ENU) Gene Knockout (ES) Genetic engineering Sterile insect technique Cloning Gene Editing Species Research Millions of Mice/Laboratory Rodents Van Eenennaam 12/8/2015 Biomedical Products Pharma Products Pigs ‐ Rabbit ‐ Ruconest Goat – ATryn, spider silk Xenotransplantation Cattle – polyclonal human antibodies Pets GloFish Micropigs Pest Control TseTse fly – sleeping sickness Mosquitoes – malaria resistance Moths – agricultural pest control Agriculture AquAdvantage Salmon – fast growth Polled Holsteins Disease resistance Improved product quality Decrease environmental footprint Single gender offspring Animal Genomics and Biotechnology Education AquAdvantage salmon Species: Atlantic salmon Breeding method: Genetic Engineering Purpose: Agriculture/Food animal Purpose Should we eat fish? Should we farm raise salmon? Does the trait impact fish health? Are there health risks from eating fast growing fish? Species Does breeding method hurt fish? Is this a food species? Do they pose an environmental hazard? Governance? Van Eenennaam 12/8/2015 Does breeding method pose new risks? Can this phenotype be achieved with other breeding methods? Has it already? Breeding Method Animal Genomics and Biotechnology Education Method Used: Selective Breeding Genomic Selection Mutagenesis (ENU) Gene Knockout (ES) Genetic engineering Sterile insect technique Cloning Gene Editing Species Photo credit: Cornell Alliance for Science Research Millions of Mice/Laboratory Rodents Van Eenennaam 12/8/2015 Biomedical Products Pharma Products Pigs ‐ Rabbit ‐ Ruconest Goat – ATryn, spider silk Xenotransplantation Cattle – polyclonal human antibodies Pets GloFish Micropigs Pest Control TseTse fly – sleeping sickness Mosquitoes – malaria resistance Moths – agricultural pest control Agriculture AquAdvantage Salmon – fast growth Polled Holsteins Disease resistance Improved product quality Decrease environmental footprint Single gender offspring Animal Genomics and Biotechnology Education Hornless Holstein Species: Bos taurus Breeding method: Gene editing Purpose: Agriculture/Food animal Photo credit: Cornell Alliance for Science Purpose Should we eat beef? Does hornless impact cattle health? Are there health risks from polled cattle? Species Does breeding method hurt cattle? Is this a food species? Do they pose an environmental hazard? Governance? Van Eenennaam 12/8/2015 Does breeding method pose new risks? Can this phenotype be achieved with other breeding methods? Has it already? Inbreeding? Breeding Method Animal Genomics and Biotechnology Education Method Used: Selective Breeding Genomic Selection Mutagenesis (ENU) Gene Knockout (ES) Genetic engineering Sterile insect technique Cloning Gene Editing Research Millions of Mice/Laboratory Rodents Van Eenennaam 12/8/2015 Biomedical Products Pharma Products Pigs ‐ Rabbit ‐ Ruconest Goat – ATryn, spider silk Xenotransplantation Cattle – polyclonal human antibodies Pets GloFish Micropigs Pest Control TseTse fly – sleeping sickness Mosquitoes – malaria resistance Moths – agricultural pest control Agriculture AquAdvantage Salmon – fast growth Polled Holsteins Disease resistance Improved product quality Decrease environmental footprint Single gender offspring Animal Genomics and Biotechnology Education Research in mice Species: Mus musculus Breeding method: ENU mutagenesis/Knockout/GE Purpose: Research Should we perform Purpose research on animals? Does breeding method impact health? Are there non‐animal substitutes to reduce experimental numbers? Species Does breeding method hurt the mice? Is this a food species? Do they pose an environmental hazard? Governance? Van Eenennaam 12/8/2015 Does breeding method pose new risks? Can this phenotype be achieved with other breeding methods? Has it already? Breeding Method Animal Genomics and Biotechnology Education Mouse x mutagenesis/genetic engineering/KO x research • There are approximately >50,000 live genetically engineered mouse lines. That is based on the fact that there are ~22000 protein coding genes in the mouse genome, and mutagenesis induced randomly (ENU) or by targeting (geneticallyengineered) elicit numerous alleles in these genes. • If you consider all the lines frozen and stored as germplasm (sperm and embryos) and embryonic stem (ES) cells from which we readily make mice, then the number of lines is far higher, likely >500,000. Kent Lloyd, UCD, Personal Communication • Thousands of off-target effects in ENU lines • According to one estimate, 10 to 50 million GE laboratory animals are used annually in the United States (Mak 2008). Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Method Used: Selective Breeding Genomic Selection Mutagenesis (ENU) Gene Knockout (ES) Genetic engineering Sterile insect technique Cloning Gene Editing Species Research Millions of Mice/Laboratory Rodents Van Eenennaam 12/8/2015 Biomedical Products Pharma Products Pigs ‐ Rabbit ‐ Ruconest Goat – ATryn, spider silk Xenotransplantation Cattle – polyclonal human antibodies Pets GloFish Micropigs Pest Control TseTse fly – sleeping sickness Mosquitoes – malaria resistance Moths – agricultural pest control Agriculture AquAdvantage Salmon – fast growth Polled Holsteins Disease resistance Improved product quality Decrease environmental footprint Single gender offspring Animal Genomics and Biotechnology Education Gene Editing x Research purpose • Possibility of reducing number of research animals used due to efficiency of gene editing as compared to genetic engineering • Less off target effects than random ENU mutagenesis • Can generate specific research lines (e.g. targeted mutations for disease models) more efficiently and less expensively • Can perform targeted gene knockout in species that are more useful as models of human disease as compared to mice • Inexpensive nature of technology opens up use to many more laboratories potentially using many more research animals Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Species x Gene Editing x Research purpose • Gene editing can be used to perform targeted edits in many more animal species as compared to ES knockouts (mostly mice) including larger-brained animals and non-human primates • May have environmental risks associated with inadequate containment of research animals from species with potential to disturb wild populations; or those specifically edited with gene drives (e.g. insects) • Governance and regulation of this technology for research purposes is ambiguous, triggered by use of an ill-defined process rather than the attributes of the animal itself, and may differ depending upon species and intended purpose of the gene edit, and even base pair size of the genetic change Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Conclusion • Need to be careful to contextualize the ethical concerns associated with a genetically modified animal application [species/method used to make the genetic modification/purpose of modification] to avoid conflating longstanding ethical issues that are not uniquely associated with the method used to make the modification • There are many ethical issues associated with the use of animals in general, the use of animals for research purposes, animal welfare, and the intentional development and use of genetically modified animals for research purposes • Those uniquely associated with gene editing to modify germ line are a subset of this set of broader issues Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education Thanks for inviting me Alison Van Eenennaam, Ph.D. Cooperative Extension Specialist Animal Biotechnology and Genomics Department of Animal Science University of California, Davis, USA [email protected] Twittter: @BioBeef http://animalscience.ucdavis.edu/animalbiotech Van Eenennaam 12/8/2015 Animal Genomics and Biotechnology Education