Production of Transgenic Rats and Mice by the Testis-Mediated Gene Transfer

Chang, Kyu‐Tae; Ikeda, Akihiro; Hayashi, Kozaburo; Furuhata, Yasufumi; Nishihara, Masugi; Ohta, Akihiko; Ogawai, Shyoso; Takahashi, Michio

doi:10.1262/jrd.45.29

Cited by 20 publications

(21 citation statements)

References 40 publications

(52 reference statements)

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“…The percentage of transgenic pups thus generated (3 out of 17 for one construct and 2 out of 36 for another one) was in the range of that obtained by DNA microinjection and comparable to that obtained in mice (Chang et al, 1999). In vivo electroporation of DNA into mouse testes has been described (Celebi et al, 2003), however, no reports on the use of this or other gene transfer techniques or vectors in rat testes have been published (Wall, 2002;Celebi et al, 2003).…”

Section: C) Sperm-mediated Transgenesissupporting

confidence: 54%

“…Transgenic rats have been generated by germline gene transfer through injection of plasmid DNA-liposome complexes into the seminiferous tubules of male rats and subsequent mating of these males with wild-type females (Chang et al, 1999). The percentage of transgenic pups thus generated (3 out of 17 for one construct and 2 out of 36 for another one) was in the range of that obtained by DNA microinjection and comparable to that obtained in mice (Chang et al, 1999).…”

Section: C) Sperm-mediated Transgenesismentioning

confidence: 82%

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Transgenic Modifications of the Rat Genome

et al. 2005

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The laboratory rat (R. norvegicus) is a very important experimental animal in several fields of biomedical research. This review describes the various techniques that have been used to generate transgenic rats: classical DNA microinjection and more recently described techniques such as lentiviral vector-mediated DNA transfer into early embryos, sperm-mediated transgenesis, embryo cloning by nuclear transfer and germline mutagenesis. It will also cover techniques associated to transgenesis such as sperm cryopreservation, embryo freezing and determination of zygosity. The availability of several technologies allowing genetic manipulation in the rat coupled to genomic data will allow biomedical research to fully benefit from the rat as an experimental animal.

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Section: C) Sperm-mediated Transgenesissupporting

confidence: 54%

Section: C) Sperm-mediated Transgenesismentioning

confidence: 82%

Transgenic Modifications of the Rat Genome

et al. 2005

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“…The fusion gene construct of mouse metallothionein-I promoter (MT)/human growth hormone receptor (hGHR) was prepared as described in the associated paper [18]. The DNA construct labeled with FITC was prepared by Sawady Technology (Tokyo) by PCR using this MT/hGHR gene as a template with primers labeled with FITC at their 5'-ends.…”

Section: Detection Of Injected Dna In Epididymal Spermatozoamentioning

confidence: 99%

“…These studies have facilitated the idea of using sperm as a vector to transfer exoge-actually carry the DNA into eggs, and the gene thus transferred is transcriptionally active [11][12][13][14][15][16][17]. In addition, as described in the associated paper [18], we have shown that DNAs injected into the testis of either rats or mice can be transferred into eggs via sperm at fertilization (testis-mediated gene transfer). The transgenes could be expressed and transmitted in the descendants, suggesting that injected DNAs were stably integrated in the genome of offspring.…”

mentioning

confidence: 95%

Possible Mechanisms for the Testis-Mediated Gene Transfer as a New Method for Producing Transgenic Animals.

Chang

Ikeda

Hayashi

et al. 1999

Journal of Reproduction and Development

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Abstract.We have recently shown that DNAs injected into the testis of either rats or mice can be transferred into eggs via sperm at fertilization and expressed and transmitted in the descendants (testis-mediated gene transfer). In the present study, we investigated the association of epididymal and ejaculated sperm with DNA injected into the testis of rats. In the first experiment, a chimeric gene of mouse metallothionein-I promoter (MT)/human growth hormone receptor (hGHR) labeled with FITC was prepared. This gene construct was mixed with cationic liposome and injected into rat testis, and 4 days later the cauda epididymis was dissected out and prepared for histological examination. A confocal microscopic observation showed that not only the head, but also the tail, of almost all spermatozoa in the epididymis was labeled with FITC. In the next experiment, 4 days after the injection of MT/hGHR gene encapsulated by liposome into the testis, the male was allowed to mate with females and spermatozoa were recovered from the uterus in the next morning. Spermatozoa were incubated in the presence or absence of deoxyribonuclease (DNase) I and then genomic DNA was extracted. The MT/hGHR gene was detected only in the spermatozoa incubated without DNase by means of PCR. These results support the notion that, in the case of our testis-mediated gene transfer, exogenous DNA injected with liposomes is not integrated into genome of the sperm and the integration occurs after fertilization.

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“…2005). A perspective modification of this technique is testis-mediated gene transfer, where plasmid DNA-liposome complexes are injected into the testis (Chang et al 1999). We can reasonably expect that with the improvement of the transgenic technology rats will expand their usefulness to the pharmaceutical industry and fundamental research even more.…”

Section: Filipiak and Saunders 2006mentioning

confidence: 99%

Beyond the Rat Models of Human Neurodegenerative Disorders

2009

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The rat is a model of choice in biomedical research for over a century. Currently, the rat presents the best "functionally" characterized mammalian model system. Despite this fact, the transgenic rats have lagged behind the transgenic mice as an experimental model of human neurodegenerative disorders. The number of transgenic rat models recapitulating key pathological hallmarks of Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, or human tauopathies is still limited. The reason is that the transgenic rats remain more difficult to produce than transgenic mice. The gene targeting technology is not yet established in rats due to the lack of truly totipotent embryonic stem cells and cloning technology. This extremely powerful technique has given the mouse a clear advantage over the rat in generation of new transgenic models. Despite these limitations, transgenic rats have greatly expanded the range of potential experimental approaches. The large size of rats permits intrathecal administration of drugs, stem cell transplantation, serial sampling of the cerebrospinal fluid, microsurgical techniques, in vivo nerve recordings, and neuroimaging procedures. Moreover, the rat is routinely employed to demonstrate therapeutic efficacy and to assess toxicity of novel therapeutic compounds in drug development. Here we suggest that the rat constitutes a slightly underestimated but perspective animal model well-suited for understanding the mechanisms and pathways underlying the human neurodegenerative disorders.

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Production of Transgenic Rats and Mice by the Testis-Mediated Gene Transfer

Cited by 20 publications

References 40 publications

Transgenic Modifications of the Rat Genome

Transgenic Modifications of the Rat Genome

Possible Mechanisms for the Testis-Mediated Gene Transfer as a New Method for Producing Transgenic Animals.

Beyond the Rat Models of Human Neurodegenerative Disorders

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