Gene Expression in Blastocysts Following Direct Injection of DNA Into Testis

Ogawa, Shyoso; Hayashi, Kozaburo; Tada, Norio; Kurihara, Takashi; Iwaya, Makoto

doi:10.1262/jrd.41.379

Cited by 23 publications

(24 citation statements)

References 7 publications

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“…Ogawa et al [20] previously reported that a large proportion of the blastocysts recovered from the oviduct of female mice mated with males injected with liposome-encapsulated fusion gene construct, which contained mouse MT promoter-neomycin resistance gene and mouse MT promoter-E. coli. β-galactosidase gene, into the testis 3 times at 4-day intervals expressed β-galactosidase activity.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, Ogawa et al [20] demonstrated that mouse blastocysts derived from eggs fertilized by spermatozoa of male mice injected with liposome-DNA complexes within the testis expressed the gene. In this study, we have prepared 3 gene constructs; human growth hormone (hGH), hGH receptor (hGHR) and mouse leptin (mOB) genes fused to the promoter region of the mouse genes for whey acidic protein (WAP), metallothionein-I (MT) and mouse mammary tumor virus (MMTV), respectively, and attempted to produce transgenic rats and mice expressing these genes adapting the testis-mediated gene transfer method.…”

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confidence: 99%

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

Chang

Ikeda

Hayashi

et al. 1999

Journal of Reproduction and Development

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Abstract. Recent reports have shown that sperm cells incubated with foreign DNA in vitro are able to transfer the DNA into eggs at fertilization. The present study examined if an injection of DNA into the testis in vivo could generate transgenic animals via sperm ejaculated. We prepared 3 gene constructs; human growth hormone (hGH), hGH receptor (hGHR) and mouse leptin (mOB) genes fused to the promoter regions of the mouse genes for whey acidic protein (WAP), metallothionein-I (MT) and mouse mammary tumor virus (MMTV), respectively. Each gene construct mixed with cationic liposome was injected into bilateral testes of male rats or mice, and the males mated with females 3 or 4 days later. A female rat mated with a male treated with MT/hGHR gene gave birth to 17 pups, 3 of which were found to carry the transgene. The expression of hGHR mRNA was demonstrated in the liver, kidney, muscle and brain after treating with zinc in drinking water. At present, the transmission of the exogenous gene to the descendants was confirmed up to the F4 generation. Three female rats mated with 3 different males injected with MMTV/mOB fusion gene produced 10, 14 and 12 pups, respectively. The genome of 2 out of these 36 pups harbored MMTVmOB gene, though expression of mOB mRNA was not detected. Two female mice were mated with 2 male mice injected with WAP/hGH gene and produced 15 and 16 pups, 3 of which incorporated the gene. The expression of hGH mRNA in the mammary gland in these mice was confirmed. These results indicate that exogenous DNA injected into the testis as a liposome-complex can be transferred into eggs via sperm and expressed in the postpartum progeny.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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

Chang

Ikeda

Hayashi

et al. 1999

Journal of Reproduction and Development

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“…Over 80% of morula-stage embryos expressed the exogenous gene, but only some postpartum progeny were foreign-DNA-positive with a high incidence of mosaicism (Yonezawa, 2001). Repeated injection with linearized plasmid DNA containing the β-galactosidase gene encapsulated with cationic liposome into the testis of adult mice via the scrotum resulted in the transmission of exogenous DNA sequences to blastocysts through fertilization (Ogawa, 1995). Sato et al (1999Sato et al ( , 2002) also attempted to transfect testicular spermatozoa with plasmid DNA by direct injection into testes to obtain transgenic animals.…”

Section: Liposomementioning

confidence: 99%

Therapeutic Potential of Gene Transfer to Testis; Myth or Reality?

Kojima¹,

Mizuno²,

Umemoto³

et al. 2011

Gene Therapy Applications

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“…On the other hand, recently, the transfer of exogenous DNA into the oocytes via spermatozoa has been reported in birds (Hasebe et al, 1998), suggesting that sperm cells can he used as a vector for carrying the gene under certain experimental condition. Ogawa et al (1995) reported, lately, that the gene injected into the mouse testis was successfully t.ransferred to the embryos obtained from normal females which were copulated with the DNA treated males. More recently, Chang d crb.…”

Section: N T Rod I~ C T Ionmentioning

confidence: 99%