Genome Editing in Large Animals |
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| Institution: | 1. AgGenetics, Nashville, TN;2. Department of Medicine, Vanderbilt University Medical Center, Nashville, TN;3. School of Agribusiness and Agriscience, Middle Tennessee State University, Murfreesboro, TN;1. Department of Animal Science, Faculty of Agriculture, Broujerd Branch, Islamic Azad University, Broujerd, Iran;2. Razi Vaccine & Serum Research Institute (Arak Branch), Arak, Iran;1. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China;2. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;3. The State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730020, China;4. Engineering Laboratory of Sheep Breeding and Reproduction Biotechnology in Gansu Province, Minqin 733300, China;1. Unidad de Producción y Sanidad Animal, Centro de Investigación y Tecnología Agroalimentaria de Aragón (CITA)- Instituto Agroalimentario de Aragón (IA2) (CITA-Universidad de Zaragoza), Zaragoza, Spain;2. ARAID, Zaragoza, Spain;3. Université de Toulouse, INRA, ENVT, GenPhySE, Castanet-Tolosan, France;4. Departamento de Mejora Genética Animal, INIA, Madrid, Spain;1. Department of Anesthesia and Critical Care, Hôpital Lariboisière, DMU Parabol, FHU Promice, APHP. Nord – Université de Paris, Paris, France;2. MΞDISIM, Inria Paris-Saclay – LMS, Ecole Polytechnique, Palaiseau, France;3. Inserm UMR-S942, Mascot, Université de Paris, Paris, France;1. Department of Bioengineering and Technology, Gauhati University, Assam, 781014, India;2. Department of Animal Biotechnology, College of Veterinary Science, Assam Agricultural University, Assam, 781022, India;3. Advanced State Biotech Hub (Assam), College of Veterinary Science, Assam Agricultural University, Assam, 781022, India;4. Division of Veterinary Microbiology and Immunology, FVSc and AH, SKUAST-Kashmir, J&K, 190006, India;5. Department of Veterinary Microbiology, College of Veterinary Science, Assam Agricultural University, Assam, 781022, India;6. Department of Biotechnology, Gauhati University, Assam, 781014, India |
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| Abstract: | Genome editing in large animals has tremendous practical applications, from more accurate models for medical research through improved animal welfare and production efficiency. Although genetic modification in large animals has a 30-year history, until recently technical issues limited its utility. The original methods—pronuclear injection and integrating viruses—were plagued with problems associated with low efficiency, silencing, poor regulation of gene expression, and variability associated with random integration. With the advent of site-specific nucleases such as TAL effector-like nucleases and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9, precision editing became possible. When used on their own, these can be used to truncate or knockout genes through nonhomologous end joining with relatively high efficiency. When used with a template containing desired gene edits, these can be used to allow insertion of any desired changes to the genome through homologous recombination with substantially lower efficiency. Consideration must be given to the issues of marker sets and off-target effects. Somatic cell nuclear transfer is most commonly used to create animals from gene-edited cells, but direct zygote injection and use of spermatogonial stem cells are alternatives under development. In developing gene editing projects, priority must be given to understanding the potential for off-target or unexpected effects of planned edits, which have been common in the past. Because of the increasing technical sophistication with which it can be accomplished, genome editing is poised to revolutionize large animal genetics, but attention must be paid to the underlying biology to maximize benefit. |
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| Keywords: | Transgenic TALEN Farm animal |
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