Science 278, 2130–2133. Biophys. 43, 355–358. doi: 10.1007/s11248-014-9832-x, Qin, Y., Chen, H., Zhang, Y., Zhu, C., Gao, B., Yin, Y., et al. doi: 10.1007/s10529-011-0667-8, Hu, S., Ni, W., Sai, W., Zi, H., Qiao, J., Wang, P., et al. 19, 962–964. In this, the udder cells from a 6-year-old Finn Dorset white sheep were injected into an unfertilized egg from a Scottish Blackface ewe, which had its nucleus removed. Since 1998, nuclear transfer protocols have evolved that make it possible to efficiently generate transgenic goats. Kids were delivered approximately 36 hours later, the majority being healthy, active, and inquisitive, with no need of extraordinary care. Four founders out of 26 (15.38%) were BLG edits, which have shown decreased expression of BLG and abolished BLG protein production in milk (Zhou et al., 2017). To study such genes, one might expect that the methods described above would work. 110, 1811–1821. To this end, transgenic pigs expressing either human thrombomodulin (hTM) or human A20 gene (hA20) on top of one or two RCAs have been recently produced to suppress both HAR and the later stage coagulatory disorders observed in experimental porcine-to-primate xenotransplantation (Petersen et al., 2009b; Oropeza et al., 2009). Theriogenology 91, 163–172. Mol. Two identical clones of a macaque monkey, Zhong Zhong and Hua Hua, were created by researchers in China and were born in late 2017. Dev. Thus, the implementation of SCNT using this approach requires less expertise, time, and cost. Genet. This strategy was applied as a genetic method of sex control and to bias the sex ratio toward females in sheep (Zhang et al., 2018b). In addition, a set of genes have also been investigated using RNAi in a variety of ovine and caprine cells. This means that the ZF domain of three or four individual fingers can recognize and bind to DNA sequences with lengths of 9 or 12 bp. Table 7 The advantages and disadvantages of genetic manipulation tools applicable to ovine and caprine genomes. (2017). DNA transposons: nature and applications in genomics. doi: 10.1038/380064a0, Chen, K., Gao, C. (2013). The authors have introduced targeting of the CF transmembrane conductance regulator (CFTR) gene and CFTR–/– as well as CFTR+/– lambs have been produced with a severe phenotype of CF pathology similar to that of humans. These tools include sperm-mediated gene transfer, viral vectors, RNA interference, recombinases, transposons, and endonucleases. Biophys. (2016). doi: 10.1016/j.smallrumres.2012.07.020, Pereyra-Bonnet, F., Fernández-Martín, R., Olivera, R., Jarazo, J., Vichera, G., Gibbons, A., et al. Sci. Received: 09 April 2019; Accepted: 17 July 2019;Published: 03 September 2019. PLoS One 12, e0186056. Science 346, 1258096. doi: 10.1126/science.1258096, Du, C., Wang, Z., Zhang, W., Fu, S., Liu, B., Liu, Z., et al. The further development of these tools has led to the generation of identical individuals by embryo splitting (Willadsen, 1979), chimeras by aggregating embryonic cells (Fehilly et al., 1984b), and even the first cloned sheep prior to the famous Dolly from undifferentiated embryonic cells (Willadsen, 1986). 49, 43–51. (2018). Development of transgenic sheep that express the visna virus envelope gene. Examples of Transgenic Animals. One of the basic strategies that uses viral vectors is the direct intramammary injection via the teat canal for the transient production of valuable proteins in milk. doi: 10.1002/mrd.20608, Zheng, H. L., Liu, X. M., Yu, Q., Xing, R. F., Yang, Z. Y., Luo, J. Dolly lived her entire life at the Roslin Institute in Midlothian. If lactase transgene is introduced and expressed in the mammary gland, milk free from lactose will be secreted. 27, 467–487. Dev. Transgenic Res. These are simple but sophisticated mechanisms derived from nature that act in prokaryotes as an adaptive immune system against phage and foreign DNA infection by the cooperation of CRISPR sequences with Cas proteins (Mojica and Montoliu, 2016). Due to their suitable size, short gestation period, and abundant secretion of milk, sheep and goats have become important model animals in agricultural, pharmaceutical, and biomedical research. doi: 10.1038/231116a0, Doudna, J. In this case, gene-editing tools are used to generate large animal embryos with genetically engineered “organ niches” by disabling a specific gene or a number of genes that are responsible for the formation of a target organ, thus allowing human PSCs to colonize the vacant niche and generate the desired organ “gest-derived organ” (Rashid et al., 2014). Sheep named Dolly was cloned by transfer of a nucleus from a mammary (Udder) cell of an adult sheep into an egg cell. ZFNs and TALENs cleave DNA with relatively similar efficiency; however, the main advantage of TALENs over ZFNs is that TALENs are easier for design and construction (Joung and Sander, 2013). Cloning 3, 31–37. Dolly, Polly and other “ollys”: likely impact of cloning technology on biomedical uses of livestock. Furthermore, Zhu et al. (2018). the gene you desire (e.g., the insulin gene). 145, 99–102. Question From class 12 Chapter BIOTECHNOLOGY: PRINCIPLE'S AND PROCESSES. Natl. doi: 10.1002/mrd.1143, Keywords: sheep, goats, farm animals, genetic modification, genome engineering, gene editing, CRISPR/Cas9, Citation: Kalds P, Zhou S, Cai B, Liu J, Wang Y, Petersen B, Sonstegard T, Wang X and Chen Y (2019) Sheep and Goat Genome Engineering: From Random Transgenesis to the CRISPR Era. To date, a number of sheep and goat models have been generated via CRISPR/Cas9 systems (Table 3). Disruption of the sheep BMPR-IB gene by CRISPR/Cas9 in in vitro-produced embryos. Breed. Furthermore, gene-edited goats that carry both BLG knockout and enriched expression of human lactoferrin (hLF) (Cui et al., 2015) or human α-lactalbumin (hLA) (Zhu et al., 2016b) have been generated using SCNT. At UNIFOR, the dam is pre-induced with 2 mg triamcinolone on day 140–142, with the induction with DEX+PGF2α on day 144–146; kids are delivered 24–30 hours later by elective C-section. Increasing the efficiencies of mediating specific genetic modifications and simplifying the procedures for generating genetically modified organisms were the main aims that challenged specialists in this field. The examined brain tissues of a transgenic cloned fetus from lentiviral-shRNA transfected fibroblasts showed a significant decrease in PrP expression (Golding et al., 2006). doi: 10.1038/nature17946, Kumar, R., Singh, S. P., Kumari, P., Mitra, A. Biomembr. Dev. Genet. New insights and current tools for genetically engineered (GE) sheep and goats. Three recipients carrying singleton fetuses reached late pregnancy, but routine ultrasound examination (once a week) indicated fetal distress in two of the fetuses close to term. Plant-produced bioscavengers have key advantages including equivalent quality and improved safety of the product (plants do not contain human pathogens or agents such as prions) as well as significant savings on production costs.
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