Genome Editing in Large Animals.
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 TALEN and CRISPR/Cas9, precision editing became possible. When used on their own, these can be used to truncate or knockout genes through non-homologous end joining (NHEJ) 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 (HR) 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 in order to maximize benefit.
Publication Date: 2016-03-25 PubMed ID: 27766006PubMed Central: PMC5067081DOI: 10.1016/j.jevs.2016.03.008Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Animal Models
- Animal Science
- Animal Studies
- Biomedical Research
- Biotechnology
- Diagnosis
- Disease Diagnosis
- Disease Management
- Disease Prevention
- Disease Treatment
- Equine Diseases
- Equine Health
- Equine Science
- Genetics
- Genomics
- Reproduction
- Reproductive Technology
- Veterinary Medicine
- Veterinary Procedure
- Veterinary Research
- Veterinary Science
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research article discusses the advancements and challenges in the process of genome editing in larger animals with the aim to increase the accuracy of medical models, enhance animal welfare and improve production efficiency.
Utility and Limitations of Genome Editing in Large Animals
- Genome editing in large animals has tremendous practical applications. It can create more accurate models for medical research, improve animal welfare, and increase production efficiency.
- Despite being a practice with a 30-year history, genome editing in large animals has faced significant limitations due to technical issues.
- Early methods, such as pronuclear injection and integrating viruses, were problematic due to low efficiency, poor regulation of gene expression, and variability linked to random integration.
Advancements in Genome Editing Techniques
- The advent of site-specific nucleases, such as TALEN and CRISPR/Cas9, has made precision genome editing possible.
- When used alone, these techniques can effectively truncate or knockout genes through a process called non-homologous end joining (NHEJ).
- When used with a template containing the desired gene changes, these methods can enable the insertion of changes to the genome through a process called homologous recombination (HR), although at a lower efficiency.
Concerns and Consideration in Genome Editing
- The article urges readers to take into consideration the issues of marker sets and potential off-target effects.
- Somatic cell nuclear transfer is the most common method utilized to create animals from gene-edited cells. Alternatives include direct zygote injection and the use of spermatogonial stem cells.
- During the development of gene editing projects, priority should be given to understanding the potential for off-target or unexpected results of planned edits, a concern that has been prominent in the past.
Future of Genome Editing in Large Animals
- Given the increasing technical sophistication, genome editing is set to revolutionize large animal genetics. However, to maximize benefits, researchers must pay close attention to the nuances of the underlying biology.
Cite This Article
APA
West J, Gill WW.
(2016).
Genome Editing in Large Animals.
J Equine Vet Sci, 41, 1-6.
https://doi.org/10.1016/j.jevs.2016.03.008 Publication
Researcher Affiliations
- AgGenetics, Nashville, TN; Department of Medicine, Vanderbilt University Medical Center, Nashville, TN.
- AgGenetics, Nashville, TN; School of Agribusiness and Agriscience, Middle Tennessee State University, Murfreesboro, TN.
Grant Funding
- R01 HL095797 / NHLBI NIH HHS
References
This article includes 44 references
- Wells DJ. Genetically modified animals and pharmacological research.. Handb Exp Pharmacol 2010;(199):213-26.
- Keiser NW, Engelhardt JF. New animal models of cystic fibrosis: what are they teaching us?. Curr Opin Pulm Med 2011 Nov;17(6):478-83.
- Houdebine LM. Production of pharmaceutical proteins by transgenic animals.. Comp Immunol Microbiol Infect Dis 2009 Mar;32(2):107-21.
- Hata T, Uemoto S, Kobayashi E. Transplantable liver production plan: "Yamaton"--liver project, Japan.. Organogenesis 2013 Oct 1;9(4):235-8.
- Phipatanakul WP, Petersen SA. Porcine small intestine submucosa xenograft augmentation in repair of massive rotator cuff tears.. Am J Orthop (Belle Mead NJ) 2009 Nov;38(11):572-5.
- Isola LM, Gordon JW. Transgenic animals: a new era in developmental biology and medicine.. Biotechnology 1991;16:3-20.
- Pursel VG, Rexroad CE Jr, Bolt DJ, Miller KF, Wall RJ, Hammer RE, Pinkert CA, Palmiter RD, Brinster RL. Progress on gene transfer in farm animals.. Vet Immunol Immunopathol 1987 Dec;17(1-4):303-12.
- Eyestone WH. Challenges and progress in the production of transgenic cattle.. Reprod Fertil Dev 1994;6(5):647-52.
- Hong SG, Kim MK, Jang G, Oh HJ, Park JE, Kang JT, Koo OJ, Kim T, Kwon MS, Koo BC, Ra JC, Kim DY, Ko C, Lee BC. Generation of red fluorescent protein transgenic dogs.. Genesis 2009 May;47(5):314-22.
- Su F, Wang Y, Liu G, Ru K, Liu X, Yu Y, Liu J, Wu Y, Quan F, Guo Z, Zhang Y. Generation of transgenic cattle expressing human β-defensin 3 as an approach to reducing susceptibility to Mycobacterium bovis infection.. FEBS J 2016 Mar;283(5):776-90.
- Hofmann A, Kessler B, Ewerling S, Kabermann A, Brem G, Wolf E, Pfeifer A. Epigenetic regulation of lentiviral transgene vectors in a large animal model.. Mol Ther 2006 Jan;13(1):59-66.
- Sugio A, Yang B, Zhu T, White FF. Two type III effector genes of Xanthomonas oryzae pv. oryzae control the induction of the host genes OsTFIIAgamma1 and OsTFX1 during bacterial blight of rice.. Proc Natl Acad Sci U S A 2007 Jun 19;104(25):10720-5.
- Boch J, Scholze H, Schornack S, Landgraf A, Hahn S, Kay S, Lahaye T, Nickstadt A, Bonas U. Breaking the code of DNA binding specificity of TAL-type III effectors.. Science 2009 Dec 11;326(5959):1509-12.
- Christian M, Cermak T, Doyle EL, Schmidt C, Zhang F, Hummel A, Bogdanove AJ, Voytas DF. Targeting DNA double-strand breaks with TAL effector nucleases.. Genetics 2010 Oct;186(2):757-61.
- Certo MT, Morgan RA. Salient Features of Endonuclease Platforms for Therapeutic Genome Editing.. Mol Ther 2016 Mar;24(3):422-9.
- Boissel S, Jarjour J, Astrakhan A, Adey A, Gouble A, Duchateau P, Shendure J, Stoddard BL, Certo MT, Baker D, Scharenberg AM. megaTALs: a rare-cleaving nuclease architecture for therapeutic genome engineering.. Nucleic Acids Res 2014 Feb;42(4):2591-601.
- Isalan M, Choo Y, Klug A. Synergy between adjacent zinc fingers in sequence-specific DNA recognition.. Proc Natl Acad Sci U S A 1997 May 27;94(11):5617-21.
- Rath D, Amlinger L, Rath A, Lundgren M. The CRISPR-Cas immune system: biology, mechanisms and applications.. Biochimie 2015 Oct;117:119-28.
- Sherkow JS. Law, history and lessons in the CRISPR patent conflict.. Nat Biotechnol 2015 Mar;33(3):256-7.
- Nemudryi AA, Valetdinova KR, Medvedev SP, Zakian SM. TALEN and CRISPR/Cas Genome Editing Systems: Tools of Discovery.. Acta Naturae 2014 Jul;6(3):19-40.
- Whitelaw CB, Sheets TP, Lillico SG, Telugu BP. Engineering large animal models of human disease.. J Pathol 2016 Jan;238(2):247-56.
- Maruyama T, Dougan SK, Truttmann MC, Bilate AM, Ingram JR, Ploegh HL. Increasing the efficiency of precise genome editing with CRISPR-Cas9 by inhibition of nonhomologous end joining.. Nat Biotechnol 2015 May;33(5):538-42.
- Meier ID, Bernreuther C, Tilling T, Neidhardt J, Wong YW, Schulze C, Streichert T, Schachner M. Short DNA sequences inserted for gene targeting can accidentally interfere with off-target gene expression.. FASEB J 2010 Jun;24(6):1714-24.
- Tanaka M, Yamaguchi S, Yamazaki Y, Kinoshita H, Kuwahara K, Nakao K, Jay PY, Noda T, Nakamura T. Somatic chromosomal translocation between Ewsr1 and Fli1 loci leads to dilated cardiomyopathy in a mouse model.. Sci Rep 2015 Jan 16;5:7826.
- Di Matteo M, Mátrai J, Belay E, Firdissa T, Vandendriessche T, Chuah MK. PiggyBac toolbox.. Methods Mol Biol 2012;859:241-54.
- Camarasa MV, Gálvez VM. Robust method for TALEN-edited correction of pF508del in patient-specific induced pluripotent stem cells.. Stem Cell Res Ther 2016 Feb 9;7:26.
- Cradick TJ, Fine EJ, Antico CJ, Bao G. CRISPR/Cas9 systems targeting β-globin and CCR5 genes have substantial off-target activity.. Nucleic Acids Res 2013 Nov;41(20):9584-92.
- Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9.. Nat Biotechnol 2016 Feb;34(2):184-191.
- Liang P, Xu Y, Zhang X, Ding C, Huang R, Zhang Z, Lv J, Xie X, Chen Y, Li Y, Sun Y, Bai Y, Songyang Z, Ma W, Zhou C, Huang J. CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes.. Protein Cell 2015 May;6(5):363-372.
- Dreyer AK, Hoffmann D, Lachmann N, Ackermann M, Steinemann D, Timm B, Siler U, Reichenbach J, Grez M, Moritz T, Schambach A, Cathomen T. TALEN-mediated functional correction of X-linked chronic granulomatous disease in patient-derived induced pluripotent stem cells.. Biomaterials 2015 Nov;69:191-200.
- Menger L, Gouble A, Marzolini MA, Pachnio A, Bergerhoff K, Henry JY, Smith J, Pule M, Moss P, Riddell SR, Quezada SA, Peggs KS. TALEN-mediated genetic inactivation of the glucocorticoid receptor in cytomegalovirus-specific T cells.. Blood 2015 Dec 24;126(26):2781-9.
- Tan W, Carlson DF, Lancto CA, Garbe JR, Webster DA, Hackett PB, Fahrenkrug SC. Efficient nonmeiotic allele introgression in livestock using custom endonucleases.. Proc Natl Acad Sci U S A 2013 Oct 8;110(41):16526-31.
- Whitworth KM, Rowland RR, Ewen CL, Trible BR, Kerrigan MA, Cino-Ozuna AG, Samuel MS, Lightner JE, McLaren DG, Mileham AJ, Wells KD, Prather RS. Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus.. Nat Biotechnol 2016 Jan;34(1):20-2.
- Wang K, Ouyang H, Xie Z, Yao C, Guo N, Li M, Jiao H, Pang D. Efficient Generation of Myostatin Mutations in Pigs Using the CRISPR/Cas9 System.. Sci Rep 2015 Nov 13;5:16623.
- Lillico SG, Proudfoot C, King TJ, Tan W, Zhang L, Mardjuki R, Paschon DE, Rebar EJ, Urnov FD, Mileham AJ, McLaren DG, Whitelaw CB. Mammalian interspecies substitution of immune modulatory alleles by genome editing.. Sci Rep 2016 Feb 22;6:21645.
- Kubota C, Yamakuchi H, Todoroki J, Mizoshita K, Tabara N, Barber M, Yang X. Six cloned calves produced from adult fibroblast cells after long-term culture.. Proc Natl Acad Sci U S A 2000 Feb 1;97(3):990-5.
- Akagi S, Matsukawa K, Takahashi S. Factors affecting the development of somatic cell nuclear transfer embryos in Cattle.. J Reprod Dev 2014;60(5):329-35.
- Lillico SG, Proudfoot C, Carlson DF, Stverakova D, Neil C, Blain C, King TJ, Ritchie WA, Tan W, Mileham AJ, McLaren DG, Fahrenkrug SC, Whitelaw CB. Live pigs produced from genome edited zygotes.. Sci Rep 2013 Oct 10;3:2847.
- Zeng W, Tang L, Bondareva A, Honaramooz A, Tanco V, Dores C, Megee S, Modelski M, Rodriguez-Sosa JR, Paczkowski M, Silva E, Wheeler M, Krisher RL, Dobrinski I. Viral transduction of male germline stem cells results in transgene transmission after germ cell transplantation in pigs.. Biol Reprod 2013 Jan;88(1):27.
- Hammer RE, Pursel VG, Rexroad CE Jr, Wall RJ, Bolt DJ, Ebert KM, Palmiter RD, Brinster RL. Production of transgenic rabbits, sheep and pigs by microinjection.. Nature 1985 Jun 20-26;315(6021):680-3.
- Pursel VG, Bolt DJ, Miller KF, Pinkert CA, Hammer RE, Palmiter RD, Brinster RL. Expression and performance in transgenic pigs.. J Reprod Fertil Suppl 1990;40:235-45.
- Jabed A, Wagner S, McCracken J, Wells DN, Laible G. Targeted microRNA expression in dairy cattle directs production of β-lactoglobulin-free, high-casein milk.. Proc Natl Acad Sci U S A 2012 Oct 16;109(42):16811-6.
- Du SJ, Gong ZY, Fletcher GL, Shears MA, King MJ, Idler DR, Hew CL. Growth enhancement in transgenic Atlantic salmon by the use of an "all fish" chimeric growth hormone gene construct.. Biotechnology (N Y) 1992 Feb;10(2):176-81.
- Zhou Q, Wang M, Yuan Y, Wang X, Fu R, Wan H, Xie M, Liu M, Guo X, Zheng Y, Feng G, Shi Q, Zhao XY, Sha J, Zhou Q. Complete Meiosis from Embryonic Stem Cell-Derived Germ Cells In Vitro.. Cell Stem Cell 2016 Mar 3;18(3):330-40.
Citations
This article has been cited 11 times.- Popova J, Bets V, Kozhevnikova E. Perspectives in Genome-Editing Techniques for Livestock.. Animals (Basel) 2023 Aug 10;13(16).
- Yan S, Zheng X, Lin Y, Li C, Liu Z, Li J, Tu Z, Zhao Y, Huang C, Chen Y, Li J, Song X, Han B, Wang W, Liang W, Lai L, Li XJ, Li S. Cas9-mediated replacement of expanded CAG repeats in a pig model of Huntington's disease.. Nat Biomed Eng 2023 May;7(5):629-646.
- Oyelami FO, Usman T, Suravajhala P, Ali N, Do DN. Emerging Roles of Noncoding RNAs in Bovine Mastitis Diseases.. Pathogens 2022 Sep 3;11(9).
- Campbell MLH. Ethics: use and misuse of assisted reproductive techniques across species.. Reprod Fertil 2021 Jul;2(3):C23-C28.
- Hay AN, Farrell K, Leeth CM, Lee K. Use of Genome Editing Techniques to Produce Transgenic Farm Animals.. Adv Exp Med Biol 2022;1354:279-297.
- Singh P, Ali SA. Impact of CRISPR-Cas9-Based Genome Engineering in Farm Animals.. Vet Sci 2021 Jun 30;8(7).
- Dasgupta I, Flotte TR, Keeler AM. CRISPR/Cas-Dependent and Nuclease-Free In Vivo Therapeutic Gene Editing.. Hum Gene Ther 2021 Mar;32(5-6):275-293.
- Walker RL, Eggel M. From Mice to Monkeys? Beyond Orthodox Approaches to the Ethics of Animal Model Choice.. Animals (Basel) 2020 Jan 1;10(1).
- de Graeff N, Jongsma KR, Johnston J, Hartley S, Bredenoord AL. The ethics of genome editing in non-human animals: a systematic review of reasons reported in the academic literature.. Philos Trans R Soc Lond B Biol Sci 2019 May 13;374(1772):20180106.
- Bhat SA, Malik AA, Ahmad SM, Shah RA, Ganai NA, Shafi SS, Shabir N. Advances in genome editing for improved animal breeding: A review.. Vet World 2017 Nov;10(11):1361-1366.
- Austin ED, West J, Loyd JE, Hemnes AR. Translational Advances in the Field of Pulmonary Hypertension Molecular Medicine of Pulmonary Arterial Hypertension. From Population Genetics to Precision Medicine and Gene Editing.. Am J Respir Crit Care Med 2017 Jan 1;195(1):23-31.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists