transgenic pig application
Production of α-1,3-galactosyltransferase null pigs by means of nuclear transfer with fibroblasts bearing loss of heterozygosity mutations. Ceftiofur resistance in Salmonella enterica serovar Heidelberg from chicken meat and humans, Canada. Since the discovery of penicillin in 1928, antibiotics have been used to treat bacterial infections in humans and animals. Potentially, pigs that express plant or microbial genes for the synthesis of nutritionally essential amino acids can permit the feeding of low-P and low-protein diets without the need for dietary supplementation with P or crystalline amino acids. Kim TK, Eberwine JH. Greene AC. Functional expression of a Delta12 fatty acid desaturase gene from spinach in transgenic pigs. [13] transferred 110 cloned embryos to four surrogate sows, resulting in the birth of one apparently normal female piglet. Much evidence shows that bacteria naturally acquire new genes (including antimicrobial-resistant genes) to survive in a new environment or host [84]. 2001;98:13090–5. [6] reported the presence of a QTL on chromosome 4 for controlling growth from birth to 70 kg, the length of the small intestine, and fat deposition in pigs (European wild boar × Large White cross). Tilapia genetically modified with human insulin grew faster than non-transgenic siblings, and could also serve as a source of islet cells for transplantation to human subjects. In this method, two domains in the inactivated Cas9 are mutated, and this protein has only a DNA binding activity, but cannot cleave DNA. Lu D, Liu S, Shang S, Wu F, Wen X, Li Z, et al. PubMed Central Article Dev Biol. Pork provides high-quality animal protein for human consumption, and is a popular food in China and many other countries, including the United States, Canada, Japan, and Europe. Insertion of a gene into, or deletion of a gene from, the animal genome may affect the function or stability of genes existing in the organisms [37]. New York: Wiley; 2014. p. 107–36. Transgenic Res. Antiviral Res. In essence, the production of transgenic animals is based on biochemical reactions, cell biology, cell culture, embryo transfer, and fetal growth and development in recipient mothers. Compared with wild-type counterparts, the transgenic pigs had greater concentrations of four ω3 polyunsaturated fatty acids: α-linolenic acid (C18:3, ω3), eicosapentaenoic acid (C20:5, ω3), docosapentaenoic acid (C22:5, ω3), and docosahexaenoic acid (C22:6, ω3). Nature. PubMed The bacteriophages are then transfected into antibiotic-resistant bacteria (e.g., Clostridium difficile), where Cas9 is guided by the guide RNA to cut the bacterial DNA at specific sites, triggering the bacteria to self-destruct (Fig. Efficient modification of the myostatin gene in porcine somatic cells and generation of knockout piglets. such as: sheep, goats, pigs, cows, rabbits, rats, mice, fish, insects, parasites and even humans have previously been used in this modification process. This can occur naturally in the birth of identical twins. Mice, however, are the most popularly tested animals in genetic modification studies. Zhang YR, Dai ZL, Wu G, Zhang R, Li N. Expression of threonine-biosynthetic genes in mammalian cells and transgenic mice. [82] used the CRISPR-Cas9 system to knock out genes responsible for AMR and re-sensitize the multidrug resistant bacteria, so that they are killed by antibiotics. Animal cells and bacteria contain the plasma membrane, cytoplasm, and nucleus, while most animal cells also contain mitochondria. 2017;546:586–7. Full text views reflects the number of PDF downloads, PDFs sent to Google Drive, Dropbox and Kindle and HTML full text views. [79] produced pigs with a functional uncoupling protein 1 (UCP1). Ames: CAST; 2008. Lai L, Kang JX, Li R, Wang J, Witt WT, Yong HY, et al. 1992;53:313–27. An important approach to solving these problems is the use of new biotechnologies, including cloning, genetic engineering (producing transgenic animals), gene editing, production of vaccines, and microbial fermentation of feedstuffs. The transgenic pig can start to hydrolyze phytates and non-starch polysaccharides in the mouth, and produce up to 24% less nitrogen and 44% less in wastes, compared with non-transgenic pigs fed the same diet. Assembling multiple xenoprotective transgenes in pigs. J Anim Sci Biotechnol. 2). Because of inadequate research, some people are concerned about the biosafety of proteins (e.g., recombinant bovine growth hormone) or potential by-products generated by the rDNA technology. For example, penicillin-resistant bacteria synthesize β-lactamase, which breaks down the β-lactam ring of penicillin to an inactive degradation product. This biotechnology is the foundation for producing transgenic animals (including pigs; see the section below) [24]. 2018;9:240. Gene transfer in pigs usually yields less than 1% transgenic piglets per injected zygote. 2013;1:203–19. Nat Biotechnol. The basic strategy of rDNA technologies is to insert a DNA fragment of interest (e.g., a porcine DNA) into a vector carrier (a DNA molecule or a plasmid) that is capable of independent replication in a host cell (e.g., E. coli). 2011;21(6):979. 12 Several laboratories have reported success with producing gene-edited pigs, which can potentially serve as organ donors, disease models, bioreactors, inactivation of porcine endogenous retrovirus in pigs, or founder animals of genetic lines with enhanced productivity (e.g., muscle growth) or disease resistance traits [59,60,61,62,63,64]. Using the SCNT technique, Betthauser et al. J Virol. Synth Syst Biotechnol. Of note, recognition of the target DNA by Cas9 is dependent on the presence of a short protospacer adjacent motif (PAM) sequence located directly downstream on the untargeted DNA strand [38]. J Adv Pharm Technol Res. When donor cells (e.g., skin cells) are from fetuses, young or mature animals, cloning is called somatic cell nuclear transfer (SCNT). Google Scholar. Application of new biotechnologies for improvements in swine nutrition and pork production. Third, swine have a suboptimal ability to digest plant-source protein, minerals and fibers. Energy supply is vital for cell integrity and function. The costs are low and benefits are enormous. Third, CRISPR/Cas9 gene targeting and SCNT technologies have been used to create pigs without the CD163 gene that encodes a cellular receptor for the porcine reproductive and respiratory syndrome virus-1 (PRRSV-1, also referred to as “blue ear disease” virus) [59]. The transformed ovum is developed into a blastocyst in vitro and the blastocyst is then transferred into a surrogate female for development to term. A trait (or characteristic) is controlled by two variant forms of a gene (called an allele) located at the same position (genetic locus) in the pair of chromosomes, with one allele inherited from each parent. 2003;5:165–80. The generation of transgenic pigs is an entirely new way of breeding. Livest Sci. Asian-Aust J Anim Sci. More work is required to address these important issues. Whitworth et al. Xenotransplantation. Generation of PPARγ mono-allelic knockout pigs via zincfinger nucleases and nuclear transfer cloning. However, these beneficial traits were offset by negative effects, including reduced reproductive performance, the onset of diseases (e.g., arthritis, gastric ulcers, dermatitis, and renal disease) and premature death [50]. Journal of Animal Science and Biotechnology [52] introduced a plant gene ∆12 fatty acid desaturase into the white adipose tissue of pigs. For example, the availability of feed-grade amino acids can substantially reduce the content of protein in diet, thereby decreasing the excretion of nitrogen into the environment. J Virology. 2014;9(4):e95097. Allan MF, Thallman RM, Cushman RA, Echternkamp SE, White SN, Kuehn LA, et al. Of note, insertion of the mouse adiponectin-UCP1 gene into the porcine endogenous UCP1 locus via the CRISPR/Cas9 as an editor can generate UCP1-knockin pigs that exhibit an improved ability to maintain body temperature, a decreased white fat mass, and an increased lean carcass yield [79]. This line of pigs had an increased ability to digest feed phytate. Furthermore, transgenic pigs [e.g., α-1,3-galactosyltransferase locus (GGTA1) knock-out swine produced through nuclear transfer by using gene-targeted fibroblasts] can provide organs for xenotransplantation in biomedicine [44]. Transgenic mice convert carbohydrates to essential fatty acids. In contrast, genome editing tools can provide more precise, more specific, more predictable and more rapid solutions to solving these problems at relatively affordable costs [38]. These short-comings can be partly overcome by the development of gene (genome) editing approach, which uses a designer nuclease (as a pair of molecular scissors) to generate a double strand break (DSB) in DNA at a desired genomic locus (Fig. Gene targeting, genome editing: from Dolly to editors. PubMed Central The transformed ovum is developed into a blastocyst in vitro and the embryo is then transferred into a surrogate mother for development to term. Some of the offspring (transgenic) contain the rDNA that has been integrated into their own genome. Bikard D. Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials. CD163 knockout pigs are fully resistant to highly pathogenic porcine reproductive and respiratory syndrome virus. Compared to the ZFN and TALEN, CRISPR/Cas9 offers greater ease of design and greater flexibility in genetic engineering, but has a higher frequency of off-target effects. Transgenic animals are those that have been genetically modified. Jazayeri SH, Amiri-Yekta A, Bahrami S, Gourabi H, Sanati MH, Khorramizadeh MR. Vector and cell line engineering technologies toward recombinant protein expression in mammalian cells lines. Mol Reprod Dev. 3). A few years later, transgenic pigs were generated that over-expressed bovine growth hormone or growth hormone releasing factor to increase their growth rate and feed efficiency, while reducing the content of body fat and plasma cholesterol levels [50, 51]. The CRISPR-Cas system has an ability to selectively target specific DNA sequences and, therefore, can easily distinguish between pathogenic or commensal bacterial species. 2018;185:986–1003. The domesticated pig has 19 pairs of chromosomes (a total of 38 chromosomes), with one set of chromosomes from each parent. 2014;32:1146–50. Polejaeva IA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, et al. GW had the primary responsibility for the content of the paper. Article Advantages and applications. London: Camco; 2012. p. 1–29. After 276 attempts, animal scientists at a Scottish institute succeeded at employing the SCNT method to produce a healthy lamb called Dolly from cells of the mammary gland of a 6-year-old sheep in 1996 [12]. In research, the RNA-Seq can be used to determine the presence and quantity of RNA (including mRNA, tRNA, and micro RNA) in a biological sample. This paper is based on the invited talk of G.W. Article Pigs lacking the scavenger receptor cysteine-rich domain 5 of CD163 are resistant to PRRSV-1 infection. Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements. Transgenic animal technology allows for the introduction of a foreign gene into the germ line of an animal to establish a desirable trait (e.g., high rates of lean tissue gain and feed efficiency) and a new capacity (e.g., synthesis of a protein with nutritional applications) in a breeding line of livestock. Nature Biotechnol. Niu D, Wei HJ, Lin L, George H, Wang T, Lee IH, et al. To date, genetically modified pigs have been generated to express bovine growth hormone, bacterial phytase, fungal carbohydrases, plant and C. elagan fatty acid desaturases, and uncoupling protein-1; and to lack myostatin, α-1,3-galactosyltransferase, or CD163 (a cellular receptor for the "blue ear disease" virus). Chung CS, Etherton TD, Wiggins JP. This revolutionary method is now used increasingly to analyze gene expression and discover SNPs in animals. Solomon MB, Pursel VG, Paroczay EW, Bolt DJ. Animals carrying these mutations should not be used for breeding.
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