Myostatin knockdown and its effect on myogenic gene expression program in stably transfected goat myoblasts. Moreover, harvesting the protein is even easier using this method (Masih et al., 2014). ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. Methods Chem. Mol. Biotechnol. Efforts in embryo manipulation research led to the development of technical tools that enabled the production of identical individuals as well as intra- and inter-specific chimeric individuals (Willadsen, 1979; Fehilly et al., 1984a; Fehilly et al., 1984b). While much of the research was done at LSU . A human lysozyme-expressing vector for injection into cattle mammary glands has also been created and reported to reduce mastitis symptoms within days (Sun et al., 2006). (2018). Biochem. Sci. The procedure of this technique is based on a combination of gene-edited embryos of large animals and human pluripotent stem cells (PSCs). Sci. |, Overview of Genetic Modification and Relevant Biotechnological Advances in Sheep and Goats, Applications of CRISPR/Cas9 in Sheep and Goats, Creative Commons Attribution License (CC BY). The prion protein is expressed mainly in the nervous system on the surface of neurones and astrocytes, but also in other organs, such as kidney and testes. 164, 87–93. Proc. J. Gen. Virol. Cleaving the target DNA can be achieved by binding and aligning two ZFN monomers to their corresponding DNA target sequences in a tail-to-tail orientation (Weinthal et al., 2010). PiggyBac transposon-mediated gene transfer in cashmere goat fetal fibroblast cells. Comparison of gene editing efficiencies of CRISPR/Cas9 and TALEN for generation of MSTN knock-out cashmere goats. This study also indicates the Rosa26 locus as a potential site for exogenous gene expression in sheep. 23, 33–40. Table 1 Examples of transgenic sheep and goats produced using a pronuclear microinjection (PNI) approach. Commun. Protein Expr. A further strategy is based on the injection of testis with shRNA vectors, which was performed to target the zinc finger protein Y-linked (ZFY) gene. In general, RNAi-based approaches are an interesting tool for mediating loss of gene expression by targeting the products of gene transcription (mRNA). 38, 177–183. doi: 10.1071/RD9890147, Nakagawa, T., Hoogenraad, C. C. (2011). Testis mediated gene transfer: in vitro transfection in goat testis by electroporation. Rev. Currently, CRISPR systems dominate the field of genome editing. doi: 10.1002/mrd.22014, Zhang, C., Wang, L., Ren, G., Li, Z., Ren, C., Zhang, T., et al. The simultaneous occurrence of FGF5 disruption at both the morphological and genetic levels has been confirmed and an enhancement in fiber length, as well as an increase in the number of secondary hair follicles were obtained (Wang et al., 2016a). Reprod. (2016). In this study, a number of both frozen and non-frozen microinjected sheep embryos have been used to compare and generate live transgenic offspring. View all Science and technology of farm animal cloning: state of the art. Expanding the genetic editing tool kit: ZFNs, TALENs, and CRISPR-Cas9. Sci. Non-viral transfection of goat germline stem cells by nucleofection results in production of transgenic sperm after germ cell transplantation. This is an example of a "transgenic" organism. (2006). Different procedures are involved in the delivery of DNA constructs as well as the various enzymes and systems that induce genetic modification events within genomes. Transgenic animals that produce useful biological products can be created by the introduction of the portion of DNA (or genes) which codes for a particular product such as human protein (a-1-antitrypsin) used to treat emphysema, tissue plasmogen activator (goat), blood clotting factors VIII and IX (sheep) and lactoferrin (cow). Advances in interspecific pregnancy. doi: 10.1126/science.6272397, Gordon, J. W., Scangos, G. A., Plotkin, D. J., Barbosa, J. Meir, Y. J. J., Wu, S. C. Y. four transgenic hLZ female goats and one non-transgenic female goat with the same breed, parity , and stage of lactation. According to the first model, the equilibrium between PrPC and PrPSc can be changed in favor of PrPSc either by a spontaneous PrPSc formation or by external PrPSc addition. Transgenic sheep designed for transplantation studies. Learn more in the Cambridge English-Chinese traditional Dictionary. J. Bioenerg. Res. doi: 10.1007/978-1-4939-8831-0_22, Fehilly, C. B., Willadsen, S. M., Tucker, E. M. (1984a). doi: 10.1016/j.pep.2007.10.015, Zhang, J., Liu, J., Yang, W., Cui, M., Dai, B., Dong, Y., et al. Animal Biology 2009. 29, 663–673. Anim. 22, 374–382. doi: 10.1073/pnas.91.15.6840, Baguisi, A., Behboodi, E., Melican, D. T., Pollock, J. S., Destrempes, M. M., Cammuso, C., et al. (2004). Biotechnol. Other clostridial endolysins that kill food pathogens have been reported (Simmons et al., 2010; Zimmer et al., 2002). (2011). The obtained results of this report support the reliability of CRISPR/Cas9 application in large animals. Currently, gene-edited sheep and goats are generated with relative ease using CRISPR systems, providing valuable models for agricultural, veterinary, and biomedical research. Goats. Sign up and get access to hundreds of high quality instruction. Commun. (2015). doi: 10.7150/ijbs.23890, Hati Boruah, J. L., Ranjan, R., Gogoi, H., Pandey, S. K., Kumar, D., Phukan, A. J., et al. It was considered different from a normal cow as it produced human protein-enriched milk. Produce 60 percent of the lysozyme and lactoferrin found in human mother's milk. A recent study in a pig model used genetic manipulation via CRISPR/Cas9 for the generation of male recipient models for SSC transplantation by targeting the nanos C2HC-type zinc finger 2 (NANOS2) gene (Park et al., 2017). Despite the relatively low efficiency of SCNT and the potential for developmental anomalies, in parallel with the PNI technique, SCNT has become a basic and dominant methodology to generate transgenic and gene-targeted sheep and goats. It is used to prevent blood clots in patients with a rare blood disease. The IgH locus, which is ~100 kb in length, is reorganized by means of the tail integration of two plasmids; this combination is constructed by segments in each cosmids, is then expressed (Masih et al., 2014). doi: 10.1139/g89-167, Williams, D. K., Pinzón, C., Huggins, S., Pryor, J. H., Falck, A., Herman, F., et al. Gene 419, 70–74. Deletion of the α (1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep. Complete answer: The first genetically modified cow or transgenic cow was Rosie. In recent years, the focus has been on the production of “transgenic cattle” (Gavin et al., 2014; Masih et al., 2014). doi: 10.1007/s12010-013-0582-7, Kumar, R., Singh, S. P., Mitra, A. 27, 467–487. Targeting human α-lactalbumin gene insertion into the goat β-lactoglobulin locus by TALEN-mediated homologous recombination. (2014). For babies of . Knockdown of endogenous myostatin promotes sheep myoblast proliferation. Ther. The PiggyBac transposon system has also been used to mediate overexpression of the thymosin β-4 (Tβ4) gene to improve the production of fine hair in cashmere goats (Shi et al., 2017). 27, 44–51. (2016b). Mammalian nuclear transfer. Nature 533, 420–424. In addition, more promising derivative tools of CRISPR systems have emerged such as base editors which enable the induction of single-base alterations without any requirements for homology-directed repair or DNA donor. Her name comes from a protein she carries in her milk, glucocerebrosidase, which works to process . PNI, cytoplasmic injection (CI), perivitelline space injection (PSI), and zona-free transduction (ZFT) have been used for the delivery to embryos, transfection (TF), and transduction (TD) for the delivery to cells, incubation (IC) for the delivery to spermatozoa, intratesticular injection (ITI) for the delivery to testes, intramammary injection (IMI) for the delivery to mammary glands, and direct injection (DI) for the delivery to targeted organs (mainly for medical purposes). Nature 231, 116–117. doi: 10.1080/10409238.2017.1304354, Ivics, Z., Hackett, P. B., Plasterk, R. H., Izsvák, Z. Alteration of sheep coat color pattern by disruption of ASIP gene via CRISPR Cas9. Bull. Natural hybridization between sheep and goats is in most cases accompanied by high mortality rates of hybrid fetuses during the second month of pregnancy, likely as a result of fetal/maternal immunological incompatibility (Dent et al., 1971). Genetically Engineered Crops builds on previous related Academies reports published between 1987 and 2010 by undertaking a retrospective examination of the purported positive and adverse effects of GE crops and to anticipate what emerging ... J. Biotechnol. Biotechnol. doi: 10.1016/S2095-3119(12)60194-4, Tan, W., Carlson, D. F., Lancto, C. A., Garbe, J. R., Webster, D. A., Hackett, P. B., et al. (2012). Another example of CRISPR/Cas9 application for the manipulation of milk components includes the generation of gene-edited sheep with an enriched production of melatonin in milk (Ma et al., 2017). Fertility and germline transmission of donor haplotype following germ cell transplantation in immunocompetent goats. The following paragraphs outline the applications of CRISPR/Cas9 in sheep and goats. doi: 10.1172/jci.insight.123529, Fan, Z., Regouski, M., Yang, M., Gash, K., Meng, Q., Davies, C. J., et al. Moreover, ear fibroblasts isolated from transgenic lambs that carried shRNA against the FMDV-VP1 gene showed a significant inhibitory effect on the VP1 gene (Deng et al., 2017). After full-term gestation, one lamb was born and died soon after birth. Efficient nonmeiotic allele introgression in livestock using custom endonucleases. Genetically modified animals are animals that have been genetically modified for a variety of purposes including producing drugs, enhancing yields, increasing resistance to disease, etc. J. Biotechnol. J. Biotechnol. Transgenic Res. If that is fails then the second approach will be to inseminate goats with sperm from the founder males of the Nexia herd. Sci. These have been employed to create targeted DSBs in eukaryotic genomes (Daboussi et al., 2015). Thus a transgenic animal that carries multiple copies of human Igs is obtained. doi: 10.1007/s11033-013-2945-0, Zhao, X., Ni, W., Chen, C., Sai, W., Qiao, J., Sheng, J., et al. In this study, a single point mutation in the tissue-nonspecific alkaline phosphatase (TNSALP) gene (ALPL, alkaline phosphatase, biomineralization associated) has been introduced. PLoS One 10, e0129457. Reprod. Theriogenology 28, 837–847. Sci. (2008b). What is produced in the milk of transgenic goats? Stanley Prusiner systematically continued working on this idea and elaborated the ‘protein-only theory’ in 1982, postulating that the misfolded protein itself was the infectious agent and designating it as proteineaceous infectious particle (‘proin,’ this later was changed to ‘prion’). -based biotechnology company. 40 knockout of goat nucleoporin 155 (NUP155) gene using CRISPR/Cas9 systems [Abstract]. Cell 135, 1017–1027. 133, 370–376. 25, 250–253. Gene editing has been revolutionized as a result of the rapid emergence of novel varieties of tools that can simply, precisely, and more efficiently mediate different forms of DNA modifications than previously reported tools. 177, 486–497. 15, 308–321. After the first generation of transgenic cloned sheep reported in 1997 (Schnieke et al., 1997), and the first transgenic cloned goats in 1999 (Baguisi et al., 1999), a large number of transgenic and gene-targeted cloned sheep and goats have been generated. (1997). J. Biotechnol. SMGT application in sheep and goats includes the production of both transgenic embryos (Pereyra-Bonnet et al., 2008; Shadanloo et al., 2010; Pereyra-Bonnet et al., 2011; Pramod et al., 2016) and transgenic founders (Zhao et al., 2010) using marker transgenes. Nontransgenic uses of PG hydrolases in food applications are limited. Somat … doi: 10.1016/0093-691X(85)90225-0. The authors wish to thank all the colleagues in Prof. Chen Yulin’s laboratory for their valuable comments during the preparation of the manuscript. CRISPR/Cas9 has also been used to investigate the functional role of genes such as stearoyl-CoA desaturase 1 (SCD1) in goat mammary epithelial cells (Tian et al., 2018a) and acetyl CoA acyltransferase 2 (ACAA2) in sheep precursor adipocyte cells (Zhang et al., 2019d). Instead of waiting for orb-producing spiders to weave a web, the technology allows for the synthesizing of up to 7 different types of silk. Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts. (2008). doi: 10.1111/febs.14520, Zhang, J., Li, L., Cai, Y., Xu, X., Chen, J., Wu, Y., et al. Nat. Due to the mosaic nature of goat and sheep tissues in the produced chimeras, chimeric characteristics cannot be transferred to the next generation. Of these tools, the four classes of site-specific endonucleases (meganucleases, ZFNs, TALENs, and CRISPRs) have attracted wide attention due to their DNA double-strand break-inducing role, which enable desired DNA modifications based on the stimulation of native cellular DNA repair mechanisms. The Flp/FRT site–specific recombination system has been applied in goat somatic cells to mediate the site-specific integration and to eliminate the problematic random integration of transgenes (Yu et al., 2013b). TALENs combined with single-stranded oligodeoxynucleotides (ssODNs) carrying a stop codon to target exon 2 of the MSTN gene have been transfected to sheep primary fibroblasts; 11.4% (4/35) of the sequenced colonies contained the desired insertion (Zhao et al., 2016). FEBS J. (2016). 2014, 281031. doi: 10.1155/2014/281031, Yuan, Y. G., Song, S. Z., Zhu, M. M., He, Z. Y., Lu, R., Zhang, T., et al.
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