Abstract
Genome editing is a powerful technology that can efficiently alter the genome of organisms to achieve targeted modification of endogenous genes and targeted integration of exogenous genes. Current genome-editing tools mainly include ZFN, TALEN and CRISPR/Cas9, which have been successfully applied to all species tested including zebrafish, humans, mice, rats, monkeys, pigs, cattle, sheep, goats and others. The application of genome editing has quickly swept through the entire biomedical field, including livestock breeding. Traditional livestock breeding is associated with rate limiting issues such as long breeding cycle and limitations of genetic resources. Genome editing tools offer solutions to these problems at affordable costs. Generation of gene-edited livestock with improved traits has proven feasible and valuable. For example, the CD163 gene-edited pig is resistant to porcine reproductive and respiratory syndrome (PRRS, also referred to as “blue ear disease”), and a SP110 gene knock-in cow less susceptible to tuberculosis. Given the high efficiency and low cost of genome editing tools, particularly CRISPR/Cas9, it is foreseeable that a significant number of genome edited livestock animals will be produced in the near future; hence it is imperative to comprehensively evaluate the pros and cons they will bring to the livestock breeding industry. Only with these considerations in mind, we will be able to fully take the advantage of the genome editing era in livestock breeding.
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Aldemita RR, Reano IM, Solis RO, Hautea RA (2015) Trends in global approvals of biotech crops (1992–2014). GM Crops Food 6:150–166
Araki M, Nojima K, Ishii T (2014) Caution required for handling genome editing technology. Trends Biotechnol 32:234–237
Burkard C, Lillico SG, Reid E, Jackson B, Mileham AJ, Ait-Ali T, Whitelaw CB, Archibald AL (2017) Precision engineering for PRRSV resistance in pigs: macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function. PLoS Pathog 13:e1006206
Cai C, Qian L, Jiang S, Sun Y, Wang Q, Ma D, Xiao G, Li B, Xie S, Gao T, Chen Y, Liu J, An X, Cui W, Li K (2017) Loss-of-function myostatin mutation increases insulin sensitivity and browning of white fat in Meishan pigs. Oncotarget 8:34911–34922
Carlson DF, Tan W, Lillico SG, Stverakova D, Proudfoot C, Christian M, Voytas DF, Long CR, Whitelaw CB, Fahrenkrug SC (2012) Efficient TALEN-mediated gene knockout in livestock. Proc Natl Acad Sci USA 109:17382–17387
Carlson DF, Lancto CA, Zang B, Kim ES, Walton M, Oldeschulte D, Seabury C, Sonstegard TS, Fahrenkrug SC (2016) Production of hornless dairy cattle from genome-edited cell lines. Nat Biotechnol 34:479–481
Chen H, Li C, Fang M, Zhu M, Li X, Zhou R, Li K, Zhao S (2009) Understanding Haemophilus parasuis infection in porcine spleen through a transcriptomics approach. BMC Genom 10:64
Christian M, Cermak T, Doyle EL, Schmidt C, Zhang F, Hummel A, Bogdanove AJ, Voytas DF (2010) Targeting DNA double-strand breaks with TAL effector nucleases. Genetics 186:757–761
Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F (2013) Multiplex genome engineering using CRISPR/Cas systems. Science 339:819–823
Davoli R, Braglia S (2007) Molecular approaches in pig breeding to improve meat quality. Brief Funct Genomic Proteomic 6:313–321
Deng S, Yu K, Zhang B, Yao Y, Liu Y, He H, Zhang H, Cui M, Fu J, Lian Z, Li N (2012) Effects of over-expression of TLR2 in transgenic goats on pathogen clearance and role of up-regulation of lysozyme secretion and infiltration of inflammatory cells. BMC Vet Res 8:196
Doyon Y, McCammon JM, Miller JC, Faraji F, Ngo C, Katibah GE, Amora R, Hocking TD, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Amacher SL (2008) Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases. Nat Biotechnol 26:702–708
Duijvesteijn N, Knol EF, Merks JW, Crooijmans RP, Groenen MA, Bovenhuis H, Harlizius B (2010) A genome-wide association study on androstenone levels in pigs reveals a cluster of candidate genes on chromosome 6. BMC Genet 11:42
Feng W, Dai Y, Mou L, Cooper DK, Shi D, Cai Z (2015) The potential of the combination of CRISPR/Cas9 and pluripotent stem cells to provide human organs from chimaeric pigs. Int J Mol Sci 16:6545–6556
Hai T, Teng F, Guo R, Li W, Zhou Q (2014) One-step generation of knockout pigs by zygote injection of CRISPR/Cas system. Cell Res 24:372–375
Hockemeyer D, Soldner F, Beard C, Gao Q, Mitalipova M, DeKelver RC, Katibah GE, Amora R, Boydston EA, Zeitler B, Meng X, Miller JC, Zhang L, Rebar EJ, Gregory PD, Urnov FD, Jaenisch R (2009) Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27:851–857
Jonas E, de Koning DJ (2015) Genomic selection needs to be carefully assessed to meet specific requirements in livestock breeding programs. Front Genet 6:49
Lai L, Kang JX, Li R, Wang J, Witt WT, Yong HY, Hao Y, Wax DM, Murphy CN, Rieke A, Samuel M, Linville ML, Korte SW, Evans RW, Starzl TE, Prather RS, Dai Y (2006) Generation of cloned transgenic pigs rich in omega-3 fatty acids. Nat Biotechnol 24:435–436
Lee K, Kwon DN, Ezashi T, Choi YJ, Park C, Ericsson AC, Brown AN, Samuel MS, Park KW, Walters EM, Kim DY, Kim JH, Franklin CL, Murphy CN, Roberts RM, Prather RS, Kim JH (2014) Engraftment of human iPS cells and allogeneic porcine cells into pigs with inactivated RAG2 and accompanying severe combined immunodeficiency. Proc Natl Acad Sci USA 111:7260–7265
Li H, Haurigot V, Doyon Y, Li T, Wong SY, Bhagwat AS, Malani N, Anguela XM, Sharma R, Ivanciu L, Murphy SL, Finn JD, Khazi FR, Zhou S, Paschon DE, Rebar EJ, Bushman FD, Gregory PD, Holmes MC, High KA (2011) In vivo genome editing restores haemostasis in a mouse model of haemophilia. Nature 475:217–221
Li X, Yang Y, Bu L, Guo X, Tang C, Song J, Fan N, Zhao B, Ouyang Z, Liu Z, Zhao Y, Yi X, Quan L, Liu S, Yang Z, Ouyang H, Chen YE, Wang Z, Lai L (2014) Rosa26-targeted swine models for stable gene over-expression and Cre-mediated lineage tracing. Cell Res 24:501–504
Li Y, Lian D, Deng S, Zhang X, Zhang J, Li W, Bai H, Wang Z, Wu H, Fu J, Han H, Feng J, Liu G, Lian L, Lian Z (2016) Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4. J Anim Sci Biotechnol 7:38
Li R, Quan S, Yan X, Biswas S, Zhang D, Shi J (2017) Molecular characterization of genetically-modified crops: challenges and strategies. Biotechnol Adv 35:302–309
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 (2013) Live pigs produced from genome edited zygotes. Sci Rep 3:2847
Lillico SG, Proudfoot C, King TJ, Tan W, Zhang L, Mardjuki R, Paschon DE, Rebar EJ, Urnov FD, Mileham AJ, McLaren DG, Whitelaw CB (2016) Mammalian interspecies substitution of immune modulatory alleles by genome editing. Sci Rep 6:21645
Liu X, Wang Y, Guo W, Chang B, Liu J, Guo Z, Quan F, Zhang Y (2013) Zinc-finger nickase-mediated insertion of the lysostaphin gene into the beta-casein locus in cloned cows. Nat Commun 4:2565
Liu X, Wang Y, Tian Y, Yu Y, Gao M, Hu G, Su F, Pan S, Luo Y, Guo Z, Quan F, Zhang Y (2014) Generation of mastitis resistance in cows by targeting human lysozyme gene to beta-casein locus using zinc-finger nucleases. Proc Biol Sci 281:20133368
Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM (2013) RNA-guided human genome engineering via Cas9. Science 339:823–826
McPherron AC, Lee SJ (1997) Double muscling in cattle due to mutations in the myostatin gene. Proc Natl Acad Sci USA 94:12457–12461
Meuwissen TH, Hayes BJ, Goddard ME (2001) Prediction of total genetic value using genome-wide dense marker maps. Genetics 157:1819–1829
Miller JC, Holmes MC, Wang J, Guschin DY, Lee YL, Rupniewski I, Beausejour CM, Waite AJ, Wang NS, Kim KA, Gregory PD, Pabo CO, Rebar EJ (2007) An improved zinc-finger nuclease architecture for highly specific genome editing. Nat Biotechnol 25:778–785
Ni W, Qiao J, Hu S, Zhao X, Regouski M, Yang M, Polejaeva IA, Chen C (2014) Efficient gene knockout in goats using CRISPR/Cas9 system. PLoS ONE 9:e106718
Nishi M, Yasue A, Nishimatu S, Nohno T, Yamaoka T, Itakura M, Moriyama K, Ohuchi H, Noji S (2002) A missense mutant myostatin causes hyperplasia without hypertrophy in the mouse muscle. Biochem Biophys Res Commun 293:247–251
Ollivier L, Sellier P (1982) Pig genetics: a review. Ann Genet Sel Anim 14:481–544
Proudfoot C, Carlson DF, Huddart R, Long CR, Pryor JH, King TJ, Lillico SG, Mileham AJ, McLaren DG, Whitelaw CB, Fahrenkrug SC (2015) Genome edited sheep and cattle. Transgenic Res 24:147–153
Qian L, Tang M, Yang J, Wang Q, Cai C, Jiang S, Li H, Jiang K, Gao P, Ma D, Chen Y, An X, Li K, Cui W (2015) Targeted mutations in myostatin by zinc-finger nucleases result in double-muscled phenotype in Meishan pigs. Sci Rep 5:14435
Ran FA, Hsu PD, Lin CY, Gootenberg JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F (2013) Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell 154:1380–1389
Ruan J, Li H, Xu K, Wu T, Wei J, Zhou R, Liu Z, Mu Y, Yang S, Ouyang H, Chen-Tsai RY, Li K (2015) Highly efficient CRISPR/Cas9-mediated transgene knockin at the H11 locus in pigs. Sci Rep 5:14253
Sato M, Miyoshi K, Nagao Y, Nishi Y, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S (2014) The combinational use of CRISPR/Cas9-based gene editing and targeted toxin technology enables efficient biallelic knockout of the alpha-1,3-galactosyltransferase gene in porcine embryonic fibroblasts. Xenotransplantation 21:291–300
Schuelke M, Wagner KR, Stolz LE, Hubner C, Riebel T, Komen W, Braun T, Tobin JF, Lee SJ (2004) Myostatin mutation associated with gross muscle hypertrophy in a child. N Engl J Med 350:2682–2688
Tang M, Zheng X, Cheng W, Jin E, Chen H, Yang S, Cui W, Li K (2011) Safety assessment of sFat-1 transgenic pigs by detecting their co-habitant microbe in intestinal tract. Transgenic Res 20:749–758
Tang M, Qian L, Jiang S, Zhang J, Song P, Chen Y, Cui W, Li K (2014) Functional and safety evaluation of transgenic pork rich in omega-3 fatty acids. Transgenic Res 23:557–571
Tao C, Yang Y, Li X, Zheng X, Ren H, Li K, Zhou R (2016) Rapid and sensitive detection of sFAT-1 transgenic pigs by visual loop-mediated isothermal amplification. Appl Biochem Biotechnol 179:938–946
Waltz E (2012) Tiptoeing around transgenics. Nat Biotechnol 30:215–217
Waltz E (2016a) Gene-edited CRISPR mushroom escapes US regulation. Nature 532:293
Waltz E (2016b) GM salmon declared fit for dinner plates. Nat Biotechnol 34:7–9
Wang K, Ouyang H, Xie Z, Yao C, Guo N, Li M, Jiao H, Pang D (2015a) Efficient generation of myostatin mutations in pigs using the CRISPR/Cas9 system. Sci Rep 5:16623
Wang X, Yu H, Lei A, Zhou J, Zeng W, Zhu H, Dong Z, Niu Y, Shi B, Cai B, Liu J, Huang S, Yan H, Zhao X, Zhou G, He X, Chen X, Yang Y, Jiang Y, Shi L, Tian X, Wang Y, Ma B, Huang X, Qu L, Chen Y (2015b) Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system. Sci Rep 5:13878
Wei J, Wagner S, Lu D, Maclean P, Carlson DF, Fahrenkrug SC, Laible G (2015) Efficient introgression of allelic variants by embryo-mediated editing of the bovine genome. Sci Rep 5:11735
Whitworth KM, Lee K, Benne JA, Beaton BP, Spate LD, Murphy SL, Samuel MS, Mao J, O’Gorman C, Walters EM, Murphy CN, Driver J, Mileham A, McLaren D, Wells KD, Prather RS (2014) Use of the CRISPR/Cas9 system to produce genetically engineered pigs from in vitro-derived oocytes and embryos. Biol Reprod 91:78
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 (2016) Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus. Nat Biotechnol 34:20–22
Whyte JJ, Zhao J, Wells KD, Samuel MS, Whitworth KM, Walters EM, Laughlin MH, Prather RS (2011) Gene targeting with zinc finger nucleases to produce cloned eGFP knockout pigs. Mol Reprod Dev 78:2
Wood RJ (1973) Robert Bakewell (1725–1795), pioneer animal breeder, and his influence on Charles Darwin. Folia Mendeliana 58:231–242
Wu H, Wang Y, Zhang Y, Yang M, Lv J, Liu J, Zhang Y (2015) TALE nickase-mediated SP110 knockin endows cattle with increased resistance to tuberculosis. Proc Natl Acad Sci USA 112:E1530–E1539
Yang D, Yang H, Li W, Zhao B, Ouyang Z, Liu Z, Zhao Y, Fan N, Song J, Tian J, Li F, Zhang J, Chang L, Pei D, Chen YE, Lai L (2011) Generation of PPARgamma mono-allelic knockout pigs via zinc-finger nucleases and nuclear transfer cloning. Cell Res 21:979–982
Yang S, Li X, Li K, Fan B, Tang Z (2014) A genome-wide scan for signatures of selection in Chinese indigenous and commercial pig breeds. BMC Genet 15:7
Yao J, Huang J, Hai T, Wang X, Qin G, Zhang H, Wu R, Cao C, Xi JJ, Yuan Z, Zhao J (2014) Efficient bi-allelic gene knockout and site-specific knock-in mediated by TALENs in pigs. Sci Rep 4:6926
Yu S, Luo J, Song Z, Ding F, Dai Y, Li N (2011) Highly efficient modification of beta-lactoglobulin (BLG) gene via zinc-finger nucleases in cattle. Cell Res 21:1638–1640
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This work was supported by The National Transgenic Project of China (2016ZX08006-001) and National Key Basic Research Program of China (2015CB943101) and Foshan University Initiative Scientific Research Program.
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Ruan, J., Xu, J., Chen-Tsai, R.Y. et al. Genome editing in livestock: Are we ready for a revolution in animal breeding industry?. Transgenic Res 26, 715–726 (2017). https://doi.org/10.1007/s11248-017-0049-7
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DOI: https://doi.org/10.1007/s11248-017-0049-7