Production of transgenic mice, rabbits and pigs by microinjection into pronuclei

Selden, Richard F.; Springman, K.; Hondele, J.; Meyer, J; Winnacker, Ernst‐Ludwig; Kräusslich, Hans‐Georg; Brem, Gottfried; Brenig, Bertram; Goodman, Howard M.; Graf, F.; Kruff, B.

doi:10.1111/j.1439-0531.1985.tb00423.x

Cited by 136 publications

(3 citation statements)

References 3 publications

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“…Because spontaneous mutant rabbits with obvious phenotypes resembling human disease phenotypes are rare and accidently discovered by experimental animal staff, it is necessary to make genetically modified rabbits according to one's own research purposes. The technology for producing transgenic (Tg) rabbits was almost concurrently reported by German (Brem et al, 1985) and US (Hammer et al, 1985) groups in 1985, but the actual use of Tg rabbit technology as an experimental tool in the field of cardiovascular diseases was not realized until 1994 when John Taylor's laboratory at the Gladstone Institute of Cardiovascular Disease in San Francisco created the first Tg rabbit expressing human hepatic lipase (Fan et al, 1994). Later on, they also produced Tg rabbits expressing human apoB-100 (Fan et al, 1995), apoE (Huang et al, 1997;Fan et al, 1998), and apoB mRNA editing protein (Yamanaka et al, 1995).…”

Section: Transgenic Rabbitsmentioning

confidence: 99%

Genetically Modified Rabbits for Cardiovascular Research

2021

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Rabbits are one of the most used experimental animals for investigating the mechanisms of human cardiovascular disease and lipid metabolism because they are phylogenetically closer to human than rodents (mice and rats). Cholesterol-fed wild-type rabbits were first used to study human atherosclerosis more than 100 years ago and are still playing an important role in cardiovascular research. Furthermore, transgenic rabbits generated by pronuclear microinjection provided another means to investigate many gene functions associated with human disease. Because of the lack of both rabbit embryonic stem cells and the genome information, for a long time, it has been a dream for scientists to obtain knockout rabbits generated by homologous recombination-based genomic manipulation as in mice. This obstacle has greatly hampered using genetically modified rabbits to disclose the molecular mechanisms of many human diseases. The advent of genome editing technologies has dramatically extended the applications of experimental animals including rabbits. In this review, we will update genetically modified rabbits, including transgenic, knock-out, and knock-in rabbits during the past decades regarding their use in cardiovascular research and point out the perspectives in future.

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Section: Transgenic Rabbitsmentioning

confidence: 99%

Genetically Modified Rabbits for Cardiovascular Research

2021

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“…Since the first genetically modified pigs were produced [37,38], easier, cheaper, and more efficient technologies have been developed to produce such animals, starting with the SCNT method and advancing now with the electroporation method. Even though electroporation is the easiest procedure, it still needs a specific instrument and involves the handling of oocytes/zygotes outside of an incubator.…”

Section: Discussionmentioning

confidence: 99%

Production of Genetically Modified Porcine Embryos via Lipofection of Zona-Pellucida-Intact Oocytes Using the CRISPR/Cas9 System

Piñeiro-Silva

Navarro-Serna

Belda-Pérez

et al. 2023

Animals

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The generation of genetically modified pigs has an important impact thanks its applications in basic research, biomedicine, and meat production. Cloning was the first technique used for this production, although easier and cheaper methods were developed, such as the microinjection, electroporation, or lipofection of oocytes and zygotes. In this study, we analyzed the production of genetically modified embryos via lipofection of zona-pellucida-intact oocytes using LipofectamineTM CRISPRMAXTM Cas9 in comparison with the electroporation method. Two factors were evaluated: (i) the increment in the concentration of the lipofectamine–ribonucleoprotein complexes (LRNPC) (5% vs. 10%) and (ii) the concentration of ribonucleoprotein within the complexes (1xRNP vs. 2xRNP). We found that the increment in the concentration of the LRNPC had a detrimental effect on embryo development and a subsequent effect on the number of mutant embryos. The 5% group had a similar mutant blastocyst rate to the electroporation method (5.52% and 6.38%, respectively, p > 0.05). The increment in the concentration of the ribonucleoprotein inside the complexes had no effect on the blastocyst rate and mutation rate, with the mutant blastocyst rate being similar in both the 1xRNP and 2xRNP lipofection groups and the electroporation group (1.75%, 3.60%, and 3.57%, respectively, p > 0.05). Here, we showed that it is possible to produce knock-out embryos via lipofection of zona-pellucida-intact porcine oocytes with similar efficiencies as with electroporation, although more optimization is needed, mainly in terms of the use of more efficient vesicles for encapsulation with different compositions.

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“…Body size in mice can be significantly increased by integrating a growth hormone (GH) transgene (Palmiter et al 1983;Brem et al 1985) leading to the concept that similar transgenes could be of great value in domesticated animals used in commercial meat production (Vize et al 1988;Bartke 2003). To-date all GH transgenes evaluated have been constructed with heterologous promoters and the resulting transgenic animals displaying accelerated growth rate, sometimes with animals achieving body weights up to 200% of wide type siblings.…”

Section: Introductionmentioning

confidence: 98%

Tissue-specific and expression of porcine growth hormone gene in BAC transgenic mice

Jia

Lillico

et al. 2010

Transgenic Res

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One of the primary goals of traditional livestock breeding is to improve growth rate and optimise body size. Growth rate can be significantly increased by integrating a growth hormone (GH) transgene under the control of a ubiquitous promoter, but while such animals do demonstrate increased growth there are also serious deleterious side-effects to the animals health. Here we report the generation and initial characterization of transgenic mice that carried a porcine BAC encoding the porcine GH gene. We show that GH expression is restricted specifically to the pituitary, is associated with elevated IGF-1 levels, and results in growth enhancement. No negative effects to the health of the transgenic animals were detected. This initial characterisation supports the use of BAC pGH transgene in livestock studies.

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Production of transgenic mice, rabbits and pigs by microinjection into pronuclei

Cited by 136 publications

References 3 publications

Genetically Modified Rabbits for Cardiovascular Research

Genetically Modified Rabbits for Cardiovascular Research

Production of Genetically Modified Porcine Embryos via Lipofection of Zona-Pellucida-Intact Oocytes Using the CRISPR/Cas9 System

Tissue-specific and expression of porcine growth hormone gene in BAC transgenic mice

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