The role of gene editing, organoids, and in vitro phenotyping in sustainable animal production
Bethany K. Redel A * , Eun Su Jeon B , Cari C. Green B and Randall S. Prather B CA
B
C
Abstract
There is a critical need for improving animal resilience, welfare, and productivity to meet the nutritional needs of the growing global population. While selective breeding has brought about tremendous improvement in livestock genetics and improving traits, it is a relatively lengthy process to integrate beneficial alleles into the herd and it is not possible to introduce variants identified in other species. Therefore, gene editing provides researchers with a tool to rapidly overcome many of these challenges. This review highlights the advances in gene editing technology, the methods used to generate gene edited livestock, and approaches that can be used to accelerate the discovery of novel alleles linked to specific traits in vitro. Additionally, the application of organoid technology is discussed, and how that linked with gene editing technology can mimic the in vivo physiology and biological functions in vitro, providing answers to important biological questions and decreasing the number of large animals needed for research. Together, these tools will enable production agriculture to be more productive and thus better able to meet the growing worldwide demand for food.
Keywords: CRISPR-Cas, direct embryo delivery, gene editing, high-throughput phenotyping, host-pathogen interactions, livestock, organoids, somatic cell nuclear transfer.
References
Adegbola S, Moore J, Sahnan K, Tozer P, Phillips R, Warusavitarne J, Faiz O, Hart A (2017) P005 Establishing a porcine model to translate anorectal stem cell organoid models to elucidate the aetiology of perianal Crohn’s fistulae. Journal of Crohn’s and Colitis 11, S81-S82.
| Crossref | Google Scholar |
Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, Chen PJ, Wilson C, Newby GA, Raguram A, Liu DR (2019) Search-and-replace genome editing without double-strand breaks or donor DNA. Nature 576, 149-157.
| Crossref | Google Scholar | PubMed |
Bevacqua RJ, Fernandez-Martín R, Savy V, Canel NG, Gismondi MI, Kues WA, Carlson DF, Fahrenkrug SC, Niemann H, Taboga OA, Ferraris S, Salamone DF (2016) Efficient edition of the bovine PRNP prion gene in somatic cells and IVF embryos using the CRISPR/Cas9 system. Theriogenology 86, 1886-1896.e1.
| Crossref | Google Scholar |
Bourdon G, Cadoret V, Charpigny G, Couturier-Tarrade A, Dalbies-Tran R, Flores M-J, Froment P, Raliou M, Reynaud K, Saint-Dizier M, Jouneau A (2021) Progress and challenges in developing organoids in farm animal species for the study of reproduction and their applications to reproductive biotechnologies. Veterinary Research 52, 42.
| Crossref | Google Scholar | PubMed |
Brazelton VA, Jr, Zarecor S, Wright DA, Wang Y, Liu J, Chen K, Yang B, Lawrence-Dill CJ (2015) A quick guide to CRISPR sgRNA design tools. GM Crops & Food 6, 266-276.
| Crossref | Google Scholar | PubMed |
Burger BT, Beaton BP, Campbell MA, Brett BT, Rohrer MS, Plummer S, Barnes D, Jiang K, Naswa S, Lange J, Ott A, Alger E, Rincon G, Rounsley S, Betthauser J, Mtango NR, Benne JA, Hammerand J, Durfee CJ, Rotolo ML, Cameron P, Lied AM, Irby MJ, Nyer DB, Fuller CK, Gradia S, Kanner SB, Park K-E, Waters J, Simpson S, Telugu BP, Salgado BC, Brandariz-Nuñez A, Rowland RRR, Culbertson M, Rice E, Cigan AM (2024) Generation of a commercial-scale founder population of porcine reproductive and respiratory syndrome virus resistant pigs using CRISPR-Cas. The CRISPR Journal 7, 12-28.
| Crossref | Google Scholar | PubMed |
Camargo LSA, Owen JR, Van Eenennaam AL, Ross PJ (2020) Efficient one-step knockout by electroporation of ribonucleoproteins into zona-intact bovine embryos. Frontiers in Genetics 11, 570069.
| Crossref | Google Scholar |
Campbell KHS, McWhir J, Ritchie WA, Wilmut I (1996) Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64-66.
| Crossref | Google Scholar | PubMed |
Carey K, Ryu J, Uh K, Lengi AJ, Clark-Deener S, Corl BA, Lee K (2019) Frequency of off-targeting in genome edited pigs produced via direct injection of the CRISPR/Cas9 system into developing embryos. BMC Biotechnology 19, 25.
| Crossref | Google Scholar | PubMed |
Carlson DF, Tan W, Lillico SG, Stverakova D, Proudfoot C, Christian M, Voytas DF, Long CR, Whitelaw CBA, Fahrenkrug SC (2012) Efficient TALEN-mediated gene knockout in livestock. Proceedings of the National Academy of Sciences of the United States of America 109, 17382-17387.
| Crossref | Google Scholar | PubMed |
Carlson DF, Lancto CA, Zang B, Kim E-S, Walton M, Oldeschulte D, Seabury C, Sonstegard TS, Fahrenkrug SC (2016) Production of hornless dairy cattle from genome-edited cell lines. Nature Biotechnology 34, 479-481.
| Crossref | Google Scholar | PubMed |
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.
| Crossref | Google Scholar | PubMed |
Derricott H, Luu L, Fong WY, Hartley CS, Johnston LJ, Armstrong SD, Randle N, Duckworth CA, Campbell BJ, Wastling JM, Coombes JL (2019) Developing a 3D intestinal epithelium model for livestock species. Cell and Tissue Research 375, 409-424.
| Crossref | Google Scholar | PubMed |
Diel DG, Lawson S, Okda F, Singrey A, Clement T, Fernandes MHV, Christopher-Hennings J, Nelson EA (2016) Porcine epidemic diarrhea virus: an overview of current virological and serological diagnostic methods. Virus Research 226, 60-70.
| Crossref | Google Scholar | PubMed |
Eiraku M, Watanabe K, Matsuo-Takasaki M, Kawada M, Yonemura S, Matsumura M, Wataya T, Nishiyama A, Muguruma K, Sasai Y (2008) Self-organized formation of polarized cortical tissues from ESCs and its active manipulation by extrinsic signals. Cell Stem Cell 3, 519-532.
| Crossref | Google Scholar | PubMed |
Fay PC, Cook CG, Wijesiriwardana N, Tore G, Comtet L, Carpentier A, Shih B, Freimanis G, Haga IR, Beard PM (2020) Madin-Darby bovine kidney (MDBK) cells are a suitable cell line for the propagation and study of the bovine poxvirus lumpy skin disease virus. Journal of Virological Methods 285, 113943.
| Crossref | Google Scholar | PubMed |
Ferrandis Vila M, Trudeau MP, Hung Y-T, Zeng Z, Urriola PE, Shurson GC, Saqui-Salces M (2018) Dietary fiber sources and non-starch polysaccharide-degrading enzymes modify mucin expression and the immune profile of the swine ileum. PLoS ONE 13, e0207196.
| Crossref | Google Scholar | PubMed |
Fu F, Li L, Shan L, Yang B, Shi H, Zhang J, Wang H, Feng L, Liu P (2017) A spike-specific whole-porcine antibody isolated from a porcine B cell that neutralizes both genogroup 1 and 2 PEDV strains. Veterinary Microbiology 205, 99-105.
| Crossref | Google Scholar | PubMed |
Garneau JE, Dupuis M-E, Villion M, Romero DA, Barrangou R, Boyaval P, Fremaux C, Horvath P, Magadán AH, Moineau S (2010) The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468, 67-71.
| Crossref | Google Scholar | PubMed |
Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551, 464-471.
| Crossref | Google Scholar | PubMed |
Giuffra E, Tuggle CK, FAANG Consortium (2019) Functional annotation of animal genomes (FAANG): current achievements and roadmap. Annual Review of Animal Biosciences 7, 65-88.
| Crossref | Google Scholar | PubMed |
Gonzalez LM, Williamson I, Piedrahita JA, Blikslager AT, Magness ST (2013) Cell lineage identification and stem cell culture in a porcine model for the study of intestinal epithelial regeneration. PLoS ONE 8, e66465.
| Crossref | Google Scholar | PubMed |
Gordon JW, Ruddle FH (1981) Integration and stable germ line transmission of genes injected into mouse pronuclei. Science 214, 1244-1246.
| Crossref | Google Scholar | PubMed |
Guo D, Zhang L, Wang X, Zheng J, Lin S (2022) Establishment methods and research progress of livestock and poultry immortalized cell lines: a review. Frontiers in Veterinary Science 9, 956357.
| Crossref | Google Scholar | PubMed |
Hammer RE, Pursel VG, Rexroad CE, Jr, Wall RJ, Bolt DJ, Ebert KM, Palmiter RD, Brinster RL (1985) Production of transgenic rabbits, sheep and pigs by microinjection. Nature 315, 680-683.
| Crossref | Google Scholar | PubMed |
Hennig SL, Owen JR, Lin JC, Young AE, Ross PJ, Van Eenennaam AL, Murray JD (2020) Evaluation of mutation rates, mosaicism and off target mutations when injecting Cas9 mRNA or protein for genome editing of bovine embryos. Scientific Reports 10, 22309.
| Crossref | Google Scholar | PubMed |
Hill KG, Curry J, DeMayo FJ, Jones-Diller K, Slapak JR, Bondioli KR (1992) Production of transgenic cattle by pronuclear injection. Theriogenology 37, 222.
| Crossref | Google Scholar |
Hirata M, Tanihara F, Wittayarat M, Hirano T, Nguyen NT, Le QA, Namula Z, Nii M, Otoi T (2019) Genome mutation after introduction of the gene editing by electroporation of Cas9 protein (GEEP) system in matured oocytes and putative zygotes. In Vitro Cellular & Developmental Biology – Animal 55, 237-242.
| Crossref | Google Scholar | PubMed |
Hwang S-U, Eun K, Kim M, Yoon JD, Cai L, Choi H, Oh D, Lee G, Kim H, Kim E, Hyun S-H (2021) Establishment of 3D neuro-organoids derived from pig embryonic stem-like cells. International Journal of Molecular Sciences 22, 2600.
| Crossref | Google Scholar | PubMed |
Jiang C, Li L, Xue M, Zhao L, Liu X, Wang W, Feng L, Liu P (2022) Long-term expanding porcine airway organoids provide insights into the pathogenesis and innate immunity of porcine respiratory coronavirus infection. Journal of Virology 96, e0073822.
| Crossref | Google Scholar | PubMed |
Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337, 816-821.
| Crossref | Google Scholar | PubMed |
Johnsson M, Hickey JM, Jungnickel MK (2024) Building in vitro tools for livestock genomics: chromosomal variation within the PK15 cell line. BMC Genomics 25, 49.
| Crossref | Google Scholar | PubMed |
Kasloff SB, Weingartl HM (2016) Swine alveolar macrophage cell model allows optimal replication of influenza A viruses regardless of their origin. Virology 490, 91-98.
| Crossref | Google Scholar | PubMed |
Kawasaki M, Goyama T, Tachibana Y, Nagao I, Ambrosini YM (2022) Farm and companion animal organoid models in translational research: a powerful tool to bridge the gap between mice and humans. Frontiers in Medical Technology 4, 895379.
| Crossref | Google Scholar | PubMed |
Khalil HA, Lei NY, Brinkley G, Scott A, Wang J, Kar UK, Jabaji ZB, Lewis M, Martín MG, Dunn JCY, Stelzner MG (2016) A novel culture system for adult porcine intestinal crypts. Cell and Tissue Research 365, 123-134.
| Crossref | Google Scholar | PubMed |
Koltes DA, Gabler NK (2016) Characterization of porcine intestinal enteroid cultures under a lipopolysaccharide challenge. Journal of Animal Science 94, 335-339.
| Crossref | Google Scholar |
Komor AC, Kim YB, Packer MS, Zuris JA, Liu DR (2016) Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 533, 420-424.
| Crossref | Google Scholar | PubMed |
Komor AC, Zhao KT, Packer MS, Gaudelli NM, Waterbury AL, Koblan LW, Kim YB, Badran AH, Liu DR (2017) Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity. Science Advances 3, eaao4774.
| Crossref | Google Scholar |
Korinek V, Barker N, Moerer P, Van Donselaar E, Huls G, Peters PJ, Clevers H (1998) Depletion of epithelial stem-cell compartments in the small intestine of mice lacking Tcf-4. Nature Genetics 19, 379-383.
| Crossref | Google Scholar | PubMed |
Kuhnert F, Davis CR, Wang H-T, Chu P, Lee M, Yuan J, Nusse R, Kuo CJ (2004) Essential requirement for Wnt signaling in proliferation of adult small intestine and colon revealed by adenoviral expression of Dickkopf-1. Proceedings of the National Academy of Sciences 101, 266-271.
| Crossref | Google Scholar |
Lamas-Toranzo I, Galiano-Cogolludo B, Cornudella-Ardiaca F, Cobos-Figueroa J, Ousinde O, Bermejo-Álvarez P (2019) Strategies to reduce genetic mosaicism following CRISPR-mediated genome edition in bovine embryos. Scientific Reports 9, 14900.
| Crossref | Google Scholar | PubMed |
Lee K, Prather RS (2013) Advancements in somatic cell nuclear transfer and future perspectives. Animal Frontiers 3, 56-61.
| Crossref | Google Scholar |
Lee K, Farrell K, Uh K (2019) Application of genome-editing systems to enhance available pig resources for agriculture and biomedicine. Reproduction, Fertility and Development 32, 40-49.
| Crossref | Google Scholar | PubMed |
Lee K, Uh K, Farrell K (2020) Current progress of genome editing in livestock. Theriogenology 150, 229-235.
| Crossref | Google Scholar | PubMed |
Li W-R, Liu C-X, Zhang X-M, Chen L, Peng X-R, He S-G, Lin J-P, Han B, Wang L-Q, Huang J-C, Liu M-J (2017) CRISPR/Cas9-mediated loss of FGF5 function increases wool staple length in sheep. The FEBS Journal 284, 2764-2773.
| Crossref | Google Scholar | PubMed |
Li J, Li J, Jr, Zhang SY, Li RX, Lin X, Mi YL, Zhang CQ (2018) Culture and characterization of chicken small intestinal crypts. Poultry Science 97, 1536-1543.
| Crossref | Google Scholar | PubMed |
Li L, Fu F, Guo S, Wang H, He X, Xue M, Yin L, Feng L, Liu P (2019) Porcine intestinal enteroids: a new model for studying enteric coronavirus porcine epidemic diarrhea virus infection and the host innate response. Journal of Virology 93, e01682-18.
| Crossref | Google Scholar |
Li Y, Yang N, Chen J, Huang X, Zhang N, Yang S, Liu G, Liu G (2020) Next-generation porcine intestinal organoids: an apical-out organoid model for swine enteric virus infection and immune response investigations. Journal of Virology 94, e01006-20.
| Crossref | Google Scholar |
Li M, Guo X, Cheng L, Zhang H, Zhou M, Zhang M, Yin Z, Guo T, Zhao L, Liu H, Liang X, Li R (2024) Porcine kidney organoids derived from naïve-like embryonic stem cells. International Journal of Molecular Sciences 25, 682.
| Crossref | Google Scholar | PubMed |
Liu Y, Li X, He S, Huang S, Li C, Chen Y, Liu Z, Huang X, Wang X (2020a) Efficient generation of mouse models with the prime editing system. Cell Discovery 6, 27.
| Crossref | Google Scholar | PubMed |
Liu M, Yu W, Jin J, Ma M, An T, Nie Y, Teng C-B (2020b) Copper promotes sheep pancreatic duct organoid growth by activation of an antioxidant protein 1-dependent MEK-ERK pathway. American Journal of Physiology-Cell Physiology 318, C806-C816.
| Crossref | Google Scholar | PubMed |
Madsen O, Rikkers RSC, Wells JM, Bergsma R, Kar SK, Taverne N, Taverne-Thiele AJ, Ellen ED, Woelders H (2024) Transcriptomic analysis of intestinal organoids, derived from pigs divergent in feed efficiency, and their response to Escherichia coli. BMC Genomics 25, 173.
| Crossref | Google Scholar | PubMed |
Mao Z, Bozzella M, Seluanov A, Gorbunova V (2008) Comparison of nonhomologous end joining and homologous recombination in human cells. DNA Repair 7, 1765-1771.
| Crossref | Google Scholar | PubMed |
Miles LA, Burga LN, Gardner EE, Bostina M, Poirier JT, Rudin CM (2017) Anthrax toxin receptor 1 is the cellular receptor for Seneca Valley virus. The Journal of Clinical Investigation 127, 2957-2967.
| Crossref | Google Scholar | PubMed |
Mitchell J, Sutton K, Elango JN, Borowska D, Perry F, Lahaye L, Santin E, Arsenault RJ, Vervelde L (2024) Chicken intestinal organoids: a novel method to measure the mode of action of feed additives. Frontiers in Immunology 15, 1368545.
| Crossref | Google Scholar | PubMed |
Norris AL, Lee SS, Greenlees KJ, Tadesse DA, Miller MF, Lombardi HA (2020) Template plasmid integration in germline genome-edited cattle. Nature Biotechnology 38, 163-164.
| Crossref | Google Scholar | PubMed |
Orkin RW, Gehron P, McGoodwin EB, Martin GR, Valentine T, Swarm R (1977) A murine tumor producing a matrix of basement membrane. The Journal of Experimental Medicine 145, 204-220.
| Crossref | Google Scholar | PubMed |
Owen JR, Hennig SL, Mcnabb BR, Mansour TA, Smith JM, Lin JC, Young AE, Trott JF, Murray JD, Delany ME, Ross PJ, Van Eenennaam AL (2021) One-step generation of a targeted knock-in calf using the CRISPR-Cas9 system in bovine zygotes. BMC Genomics 22, 118.
| Crossref | Google Scholar | PubMed |
Palmiter RD, Brinster RL, Hammer RE, Trumbauer ME, Rosenfeld MG, Birnberg NC, Evans RM (1982) Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes. Nature 300, 611-615.
| Crossref | Google Scholar | PubMed |
Perisse IV, Fan Z, Singina GN, White KL, Polejaeva IA (2021) Improvements in gene editing technology boost its applications in livestock. Frontiers in Genetics 11, 614688.
| Crossref | Google Scholar |
Petri K, Zhang W, Ma J, Schmidts A, Lee H, Horng JE, Kim DY, Kurt IC, Clement K, Hsu JY, Pinello L, Maus MV, Joung JK, Yeh J-RJ (2022) CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells. Nature Biotechnology 40, 189-193.
| Crossref | Google Scholar | PubMed |
Pierzchalska M, Panek M, Czyrnek M, Gielicz A, Mickowska B, Grabacka M (2017) Probiotic Lactobacillus acidophilus bacteria or synthetic TLR2 agonist boost the growth of chicken embryo intestinal organoids in cultures comprising epithelial cells and myofibroblasts. Comparative Immunology, Microbiology and Infectious Diseases 53, 7-18.
| Crossref | Google Scholar | PubMed |
Pinto D, Gregorieff A, Begthel H, Clevers H (2003) Canonical Wnt signals are essential for homeostasis of the intestinal epithelium. Genes & Development 17, 1709-1713.
| Crossref | Google Scholar | PubMed |
Polejaeva IA (2021) 25th anniversary of cloning by somatic cell nuclear transfer: generation of genetically engineered livestock using somatic cell nuclear transfer. Reproduction 162, F11-F22.
| Crossref | Google Scholar | PubMed |
Powell RH, Behnke MS (2017) WRN conditioned media is sufficient for in vitro propagation of intestinal organoids from large farm and small companion animals. Biology Open 6, 698-705.
| Crossref | Google Scholar | PubMed |
Quah PS, Tran BM, Corbin VDA, Chang JJ-Y, Wong CY, Diaz-Méndez A, Hartley CA, Zeng W, Hanssen E, Trifunovic Z, Reading PC, Jackson DC, Vincan E, Coin LJM, Deliyannis G (2023) Development of matrix-embedded bovine tracheal organoids to study the innate immune response against bovine respiratory disease. Organoids 2, 82-101.
| Crossref | Google Scholar |
Redel BK, Prather RS (2016) Meganucleases revolutionize the production of genetically engineered pigs for the study of human diseases. Toxicologic Pathology 44, 428-433.
| Crossref | Google Scholar | PubMed |
Redel BK, Yoon J, Reese E, An H, Uh K, Chen PR, Prather RS, Lee K (2024) Novel off-targeting events identified after genome wide analysis of CRISPR-Cas edited pigs. The CRISPR Journal 7, 141-149.
| Crossref | Google Scholar | PubMed |
Rodríguez-Rodríguez DR, Ramírez-Solís R, Garza-Elizondo MA, Garza-Rodríguez MDL, Barrera-Saldaña HA (2019) Genome editing: a perspective on the application of CRISPR/Cas9 to study human diseases (Review). International Journal of Molecular Medicine 43, 1559-1574.
| Crossref | Google Scholar | PubMed |
Ryu J, Prather RS, Lee K (2018) Use of gene-editing technology to introduce targeted modifications in pigs. Journal of Animal Science and Biotechnology 9, 5.
| Crossref | Google Scholar | PubMed |
Sakib S, Uchida A, Valenzuela-Leon P, Yu Y, Valli-Pulaski H, Orwig K, Ungrin M, Dobrinski I (2019) Formation of organotypic testicular organoids in microwell culture. Biology of Reproduction 100, 1648-1660.
| Crossref | Google Scholar | PubMed |
Sander JD, Joung JK (2014) CRISPR-Cas systems for editing, regulating and targeting genomes. Nature Biotechnology 32, 347-355.
| Crossref | Google Scholar | PubMed |
Sato T, Vries RG, Snippert HJ, Van De Wetering M, Barker N, Stange DE, Van Es JH, Abo A, Kujala P, Peters PJ, Clevers H (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262-265.
| Crossref | Google Scholar | PubMed |
Schwarz JS, De Jonge HR, Forrest JN, Jr (2015) Value of organoids from comparative epithelia models. The Yale Journal of Biology and Medicine 88, 367-374.
| Google Scholar | PubMed |
Sharbati J, Hanisch C, Pieper R, Einspanier R, Sharbati S (2015) Small molecule and RNAi induced phenotype transition of expanded and primary colonic epithelial cells. Scientific Reports 5, 12681.
| Crossref | Google Scholar | PubMed |
Sheets T, Park C-H, Park K-E, Powell A, Donovan D, Telugu B (2016) Somatic cell nuclear transfer followed by CRIPSR/Cas9 microinjection results in highly efficient genome editing in cloned pigs. International Journal of Molecular Sciences 17, 2031.
| Crossref | Google Scholar |
Sheltzer JM, Torres EM, Dunham MJ, Amon A (2012) Transcriptional consequences of aneuploidy. Proceedings of the National Academy of Sciences of the United States of America 109, 12644-12649.
| Crossref | Google Scholar | PubMed |
Sidik SM, Huet D, Ganesan SM, Huynh M-H, Wang T, Nasamu AS, Thiru P, Saeij JPJ, Carruthers VB, Niles JC, Lourido S (2016) A genome-wide CRISPR screen in toxoplasma identifies essential apicomplexan genes. Cell 166, 1423-1435.e12.
| Crossref | Google Scholar |
Tanihara F, Takemoto T, Kitagawa E, Rao S, Do LTK, Onishi A, Yamashita Y, Kosugi C, Suzuki H, Sembon S, Suzuki S, Nakai M, Hashimoto M, Yasue A, Matsuhisa M, Noji S, Fujimura T, Fuchimoto D-I, Otoi T (2016) Somatic cell reprogramming-free generation of genetically modified pigs. Science Advances 2, e1600803.
| Crossref | Google Scholar | PubMed |
van Der Hee B, Loonen LMP, Taverne N, Taverne-Thiele JJ, Smidt H, Wells JM (2018) Optimized procedures for generating an enhanced, near physiological 2D culture system from porcine intestinal organoids. Stem Cell Research 28, 165-171.
| Crossref | Google Scholar | PubMed |
Vilarino M, Suchy FP, Rashid ST, Lindsay H, Reyes J, Mcnabb BR, Van Der Meulen T, Huising MO, Nakauchi H, Ross PJ (2018) Mosaicism diminishes the value of pre-implantation embryo biopsies for detecting CRISPR/Cas9 induced mutations in sheep. Transgenic Research 27, 525-537.
| Crossref | Google Scholar | PubMed |
von Furstenberg RJ, Li J, Stolarchuk C, Feder R, Campbell A, Kruger L, Gonzalez LM, Blikslager AT, Cardona DM, Mccall SJ, Henning SJ, Garman KS (2017) Porcine esophageal submucosal gland culture model shows capacity for proliferation and differentiation. Cellular and Molecular Gastroenterology and Hepatology 4, 385-404.
| Crossref | Google Scholar | PubMed |
Wall RJ (1996) Transgenic livestock: progress and prospects for the future. Theriogenology 45, 57-68.
| Crossref | Google Scholar |
Whitworth KM, Prather RS (2010) Somatic cell nuclear transfer efficiency: how can it be improved through nuclear remodeling and reprogramming? Molecular Reproduction and Development 77, 1001-1015.
| Crossref | Google Scholar | PubMed |
Whitworth KM, Rowland RRR, 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. Nature Biotechnology 34, 20-22.
| Crossref | Google Scholar | PubMed |
Whitworth KM, Benne JA, Spate LD, Murphy SL, Samuel MS, Murphy CN, Richt JA, Walters E, Prather RS, Wells KD (2017) Zygote injection of CRISPR/Cas9 RNA successfully modifies the target gene without delaying blastocyst development or altering the sex ratio in pigs. Transgenic Research 26, 97-107.
| Crossref | Google Scholar | PubMed |
Whitworth KM, Green JA, Redel BK, Geisert RD, Lee K, Telugu BP, Wells KD, Prather RS (2022) Improvements in pig agriculture through gene editing. CABI Agriculture and Bioscience 3, 41.
| Crossref | Google Scholar | PubMed |
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. Molecular Reproduction and Development 78, 2.
| Crossref | Google Scholar | PubMed |
Wiedenheft B, Sternberg SH, Doudna JA (2012) RNA-guided genetic silencing systems in bacteria and archaea. Nature 482, 331-338.
| Crossref | Google Scholar | PubMed |
Workman AM, Heaton MP, Vander Ley BL, Webster DA, Sherry L, Bostrom JR, Larson S, Kalbfleisch TS, Harhay GP, Jobman EE, Carlson DF, Sonstegard TS (2023) First gene-edited calf with reduced susceptibility to a major viral pathogen. PNAS Nexus 2, pgad125.
| Crossref | Google Scholar |
World Food Programme (2024) Global operational response plan #11. Available at https://www.wfp.org/ending-hunger [Accessed June 2024]
World Organisation for Animal Health (2024) Animal Health Welfare. Available at https://www.woah.org/en/what-we-do/animal-health-and-welfare/ [Accessed 22 February 2024]
Yang H, Wu Z (2018) Genome editing of pigs for agriculture and biomedicine. Frontiers in Genetics 9, 360.
| Crossref | Google Scholar | PubMed |
Yang X, Wu X, Wang Y, Li W, Wu X, Yuan L, Yu T, Li N, Zhang S, Hua J (2024) Induction of lung progenitor cell-like organoids by porcine pluripotent stem cells. The FASEB Journal 38, e23481.
| Crossref | Google Scholar | PubMed |
Zarei K, Stroik MR, Gansemer ND, Thurman AL, Ostedgaard LS, Ernst SE, Thornell IM, Powers LS, Pezzulo AA, Meyerholz DK, Stoltz DA (2020) Early pathogenesis of cystic fibrosis gallbladder disease in a porcine model. Laboratory Investigation 100, 1388-1399.
| Crossref | Google Scholar | PubMed |
Zhang Q, Ke H, Blikslager A, Fujita T, Yoo D (2018) Type III interferon restriction by porcine epidemic diarrhea virus and the role of viral protein nsp1 in IRF1 signaling. Journal of Virology 92 e01677-17.
| Crossref | Google Scholar |
Zhao Z, Chen X, Dowbaj AM, Sljukic A, Bratlie K, Lin L, Fong ELS, Balachander GM, Chen Z, Soragni A, Huch M, Zeng YA, Wang Q, Yu H (2022) Organoids. Nature Reviews Methods Primers 2, 94.
| Crossref | Google Scholar | PubMed |
Zhou S, Lenk LJ, Gao Y, Wang Y, Zhao X, Pan M, Huang S, Sun K, Kalds P, Luo Q, Lillico S, Sonstegard T, Scholl UI, Ma B, Petersen B, Chen Y, Wang X (2023) Generation of sheep with defined FecBB and TBXT mutations and porcine blastocysts with KCNJ5G151R/+ mutation using prime editing. BMC Genomics 24, 313.
| Crossref | Google Scholar | PubMed |
Zhu M, Qin Y-C, Gao C-Q, Yan H-C, Li X-G, Wang X-Q (2019) Extracellular glutamate-induced mTORC1 activation via the IR/IRS/PI3K/Akt Pathway enhances the expansion of porcine intestinal stem cells. Journal of Agricultural and Food Chemistry 67, 9510-9521.
| Crossref | Google Scholar | PubMed |
Zilova L, Weinhardt V, Tavhelidse T, Schlagheck C, Thumberger T, Wittbrodt J (2021) Fish primary embryonic pluripotent cells assemble into retinal tissue mirroring in vivo early eye development. eLife 10, e66998.
| Crossref | Google Scholar | PubMed |