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Review
. 2018 Jan 22;11(1):dmm030783.
doi: 10.1242/dmm.030783.

Genetically engineered pigs as models for human disease

Affiliations
Review

Genetically engineered pigs as models for human disease

Carolin Perleberg et al. Dis Model Mech. .

Abstract

Genetically modified animals are vital for gaining a proper understanding of disease mechanisms. Mice have long been the mainstay of basic research into a wide variety of diseases but are not always the most suitable means of translating basic knowledge into clinical application. The shortcomings of rodent preclinical studies are widely recognised, and regulatory agencies around the world now require preclinical trial data from nonrodent species. Pigs are well suited to biomedical research, sharing many similarities with humans, including body size, anatomical features, physiology and pathophysiology, and they already play an important role in translational studies. This role is set to increase as advanced genetic techniques simplify the generation of pigs with precisely tailored modifications designed to replicate lesions responsible for human disease. This article provides an overview of the most promising and clinically relevant genetically modified porcine models of human disease for translational biomedical research, including cardiovascular diseases, cancers, diabetes mellitus, Alzheimer's disease, cystic fibrosis and Duchenne muscular dystrophy. We briefly summarise the technologies involved and consider the future impact of recent technical advances.

Keywords: Disease models; Genetic modification; Pig; Swine.

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Conflict of interest statement

Competing interestsThe authors declare no competing or financial interests.

Figures

Fig. 1.
Fig. 1.
Methods used to generate genetically modified pigs. (A) Pronuclear microinjection of DNA results in random integration of transgenes into the genome, but does not enable gene targeted modifications. Viral vectors can also be microinjected to increase the frequency of transgenesis. (B) Somatic primary cells can be cultured and genetically modified by various methods to add random transgenes or for gene targeting. A genetically modified cell (shown in green) is introduced into the perivitelline space of an enucleated oocyte and an electrical pulse used to fuse the cell membranes and simultaneously activate the oocyte. (C) Endonuclease RNA or protein and guide RNA(s) are injected into the cytoplasm of the fertilised oocyte or zygote to directly modify the embryo genome.
Fig. 2.
Fig. 2.
Porcine oocytes. Note the opacity of the ooplasm caused by their high lipid content. Porcine oocytes require centrifugation to visualise their pronuclei for microinjection.
Fig. 3.
Fig. 3.
Gene editing by CRISPR/Cas9 for gene inactivation and targeted sequence replacement. During gene editing by CRISPR/Cas 9, the endonuclease Cas9 (green) is led by the guide RNA (gRNA) to the genomic target site, where it cleaves the double-stranded DNA (dsDNA) at a point 3-5 bp upstream of the protospacer adjacent motif (PAM). The resulting double-strand break (DSB) can then be repaired by nonhomologous end joining (NHEJ, left) or by homology-directed repair (HDR, right). NHEJ is an error-prone mechanism that can lead to sequence deletion, insertion or both, which can disrupt gene function. The HDR pathway is more precise and uses template DNA to repair the DSB via homologous recombination. The introduction of an exogenous DNA template, as dsDNA or as single-stranded oligodeoxynucleotide (ssODN), allows desired sequence changes to be engineered.

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