Production of transgenic pig as an Alzheimer’s disease model using a multi-cistronic vector system

Lipiński

et al. 2020

Genes

Progress in genetic engineering over the past few decades has made it possible to develop methods that have led to the production of transgenic animals. The development of transgenesis has created new directions in research and possibilities for its practical application. Generating transgenic animal species is not only aimed towards accelerating traditional breeding programs and improving animal health and the quality of animal products for consumption but can also be used in biomedicine. Animal studies are conducted to develop models used in gene function and regulation research and the genetic determinants of certain human diseases. Another direction of research, described in this review, focuses on the use of transgenic animals as a source of high-quality biopharmaceuticals, such as recombinant proteins. The further aspect discussed is the use of genetically modified animals as a source of cells, tissues, and organs for transplantation into human recipients, i.e., xenotransplantation. Numerous studies have shown that the pig (Sus scrofa domestica) is the most suitable species both as a research model for human diseases and as an optimal organ donor for xenotransplantation. Short pregnancy, short generation interval, and high litter size make the production of transgenic pigs less time-consuming in comparison with other livestock species This review describes genetically modified pigs used for biomedical research and the future challenges and perspectives for the use of the swine animal models.

Section: Alzheimer's Diseasementioning

confidence: 60%

Application of Genetically Engineered Pigs in Biomedical Research

Lipiński

et al. 2020

Genes

“…Still, when four 8-year-old and a single 14-year-old sheep were analyzed, Tau tangle-like structures were identified in 3 of the four 8-year-old sheep, and Aβ-immunopositive plaques were identified in all animals, although most plaques were diffuse 34 . AD transgenic pigs have recently been generated 53 , but the thickness of the pig skull continues to increase with aging 54 , making this species less suitable for aged, AD, or longitudinal studies. Minipigs also possess large, air-filled sinuses atop the skull, which would block ultrasound transmission.…”

Section: Discussionmentioning

confidence: 99%

Establishing sheep as an experimental species to validate ultrasound-mediated blood-brain barrier opening for potential therapeutic interventions

Pelekanos¹,

Leinenga²,

Odabaee³

et al. 2018

Theranostics

Rationale: Treating diseases of the brain such as Alzheimer's disease (AD) is challenging as the blood-brain barrier (BBB) effectively restricts access of a large number of potentially useful drugs. A potential solution to this problem is presented by therapeutic ultrasound, a novel treatment modality that can achieve transient BBB opening in species including rodents, facilitated by biologically inert microbubbles that are routinely used in a clinical setting for contrast enhancement. However, in translating rodent studies to the human brain, the presence of a thick cancellous skull that both absorbs and distorts ultrasound presents a challenge. A larger animal model that is more similar to humans is therefore required in order to establish a suitable protocol and to test devices. Here we investigated whether sheep provide such a model.Methods: In a stepwise manner, we used a total of 12 sheep to establish a sonication protocol using a spherically focused transducer. This was assisted by ex vivo simulations based on CT scans to establish suitable sonication parameters. BBB opening was assessed by Evans blue staining and a range of histological tests.Results: Here we demonstrate noninvasive microbubble-mediated BBB opening through the intact sheep skull. Our non-recovery protocol allowed for BBB opening at the base of the brain, and in areas relevant for AD, including the cortex and hippocampus. Linear time-shift invariant analysis and finite element analysis simulations were used to optimize the position of the transducer and to predict the acoustic pressure and location of the focus.Conclusion: Our study establishes sheep as a novel animal model for ultrasound-mediated BBB opening and highlights opportunities and challenges in using this model. Moreover, as sheep develop an AD-like pathology with aging, they represent a large animal model that could potentially complement the use of non-human primates.

Reprod Domestic Animals

“…In addition, pigs, especially minipigs, also share a number of similarities with human, such as body size, anatomy, physiology, pathology as well as genome, providing an excellent animal model for mimicking human diseases (Nagashima et al, ; Niemann & Lucas‐Hahn, ; Prather et al, ; Whyte & Prather, ). Genetically modified pigs have also been widely used for modelling numerous human diseases, such as Alzheimer’s disease (Kragh et al, ; Lee et al, ), Huntington’s disease (Yan et al, ; Yang et al, ) and Parkinson’s disease (Wang et al, ; Zhou et al, ). Therefore, genetically modified pigs hold great promise in the fields of agriculture and biomedicine.…”

Section: Discussionmentioning

confidence: 99%

Generation of transgenic‐cloned Huanjiang Xiang pigs systemically expressing enhanced green fluorescent protein

Zhu

Zhong²,

Ge³

et al. 2018

Huanjiang Xiang pig is a unique native minipig breed originating in Guangxi, China, and has great utility value in agriculture and biomedicine. Reproductive biotechnologies such as somatic cell nuclear transfer (SCNT) and SCNT-mediated genetic modification show great potential value in genetic preservation and utilization of Huanjiang Xiang pigs. Our previous work has successfully produced cloned and transgenic-cloned embryos using somatic cells from a Huanjiang Xiang pig. In this study, we firstly report the generation of transgenic-cloned Huanjiang Xiang pigs carrying an enhanced green fluorescent protein (eGFP) gene. A total of 504 SCNT-derived embryos were transferred to two surrogate recipients, one of which became pregnant and gave birth to three live piglets. Exogenous eGFP transgene had integrated in all of the three Huanjiang Xiang piglets identified by genotyping. Furthermore, expression of eGFP was also detected from in vitro cultured skin fibroblast cells and various organs or tissues from positive transgenic-cloned Huanjiang Xiang pigs. The present work provides a practical method to preserve this unique genetic resource and also lays a foundation for genetic modification of Huanjiang Xiang pigs with improved values in agriculture and biomedicine.