Transgenic Rabbit Models: Now and the Future

Abstract

Transgenic rabbits have contributed to the progress of biomedical science as human disease models because of their unique features, such as the lipid metabolism system similar to humans and medium body size that facilitates handling and experimental manipulation. In fact, many useful transgenic rabbits have been generated and used in research fields such as lipid metabolism and atherosclerosis, cardiac failure, immunology, and oncogenesis. However, there have been long-term problems, namely that the transgenic efficiency when using pronuclear microinjection is low compared with transgenic mice and production of knockout rabbits is impossible owing to the lack of embryonic stem cells for gene targeting in rabbits. Despite these limitations, the emergence of novel genome editing technology has changed the production of genetically modified animals including the rabbit. We are finally able to produce both transgenic and knockout rabbit models to analyze gain- and loss-of-functions of specific genes. It is expected that the use of genetically modified rabbits will extend to various research fields. In this review, we describe the unique features of rabbits as laboratory animals, the current status of their development and use, and future perspectives of transgenic rabbit models for human diseases.

Strategies for Highly Efficient Rabbit Sperm Cryopreservation

“…Along with the development of genetic engineering, a number of gene-modified rabbits have been established as experimental models of human diseases. In addition to transgenic rabbits produced with the conventional pronuclear microinjection technique, knockout rabbits have been established using CRISPR/Cas9 genome editing technology [ 4 ]. These established genetically modified rabbits are rare and valuable and thus it is vitally important to breed and maintain rabbit strains for different purposes and preserve them as bio-resources [ 5 ].…”

Transgenic rabbit models for cardiac disease research

“…Although these new techniques have already been used to knock-out genes in the field of lipid metabolism and vascular/atherosclerosis research (reviewed in (Matsuhisa et al, 2020), they have not been used for the generation of transgenic rabbit models for monogenic cardiac diseases as these need more complex approaches to introduce targeted disease-specific point mutations. However, with further modification and improvement of this technique, the insertion of targeted point mutations will also be feasible rather than only knockout or knock-in approaches of complete genes, thus allowing the future CRISPR/Cas9 based generation of transgenic rabbit models also for monogenic cardiac diseases.…”

“…Although the initial focus has been on animal models of cardiac disease induced by drugs, surgical interventions or other environmental changes (Hasenfuss, 1998), more and more genetically modified animal models have been generated in recent years (Cesarovic et al, 2020b;Matsuhisa et al, 2020). In these models, genes of interest are knocked-out or knocked-in or are subjected to disease-specific point mutations.…”

“…In the past decades, the laboratory rabbit (Oryctolagus cuniculus) has gained more and more significance in cardiac research as a useful alternative model to small rodents and larger animal models. Thanks to continued improvements in animal transgenesis (Bosze et al, 2016;Matsuhisa et al, 2020), rabbits have entered the range of species in whom genetic manipulation can successfully replicate human cardiac diseases.…”

“…Despite several attempts to improve the efficiency of microinjectionmediated transgenic animal production (Houdebine, 2002), one of the disadvantages of the pronuclear injection-based transgenesis could not be solved, namely random integration, this means that the pronuclear microinjection technology is unable to control the copy number and integration site of the transgene in the genome. Therefore, there is a high chance of a position effect, which might lead to undesired ectopic expression or transgene silencing (Ivics et al, 2014;Matsuhisa et al, 2020).…”