Sperm-mediated DNA transfer to cells of the uterus and embryo

Chan, Philip J.

doi:10.1002/(sici)1098-2795(200006)56:2+<316::aid-mrd23>3.0.co;2-v

Cited by 7 publications

(2 citation statements)

References 40 publications

(47 reference statements)

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“…These results are in accordance with previous reports that confirmed that bovine and porcine spermatozoa can bind to exogenous DNA and maintain their motility (Rieth et al 2000, Alderson et al 2006, Kang et al 2008. Interestingly, one previous report (Chan 2000) described how transfected sperm had even higher percentages of total motility and progression when compared with controls. However, some controversial aspects exist in relation to sperm motility, so in another report, transfected spermatozoa were reported to be immotile (Anzar & Buhr 2006).…”

Section: F a García-vázquez And Otherssupporting

confidence: 93%

Production of transgenic piglets using ICSI–sperm-mediated gene transfer in combination with recombinase RecA

García–Vázquez¹,

Ruiz²,

Matás³

et al. 2010

Reproduction

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Sperm-mediated gene transfer (SMGT) is a method for the production of transgenic animals based on the intrinsic ability of sperm cells to bind and internalize exogenous DNA molecules and to transfer them into the oocyte at fertilization. Recombinase-A (RecA) proteincoated exogenous DNA has been used previously in pronuclear injection systems increasing integration into goat and pig genomes. However, there are no data regarding transgene expression after ICSI. Here, we set out to investigate whether the expression of transgenic DNA in porcine embryos is improved by recombinase-mediated DNA transfer and if it is possible to generate transgenic animals using this methodology. Different factors which could affect the performance of this transgenic methodology were analyzed by studying 1) the effect of the presence of exogenous DNA and RecA protein on boar sperm functionality; 2) the effect of recombinase RecA on in vitro enhanced green fluorescent protein (EGFP)-expressing embryos produced by ICSI or IVF; and 3) the efficiency of generation of transgenic piglets by RecA-mediated ICSI. Our results suggested that 1) the presence of exogenous DNA and RecA-DNA complexes at 5 mg/ml did not affect sperm functionality in terms of motility, viability, membrane lipid disorder, or reactive oxygen species generation; 2) EGFP-expressing embryos were obtained with a high efficiency using the SMGT-ICSI technique in combination with recombinase; however, the use of IVF system did not result in any fluorescent embryos; and 3) transgenic piglets were produced by this methodology. To our knowledge, this is the first time that transgenic pigs have been produced by ICSI-SGMT and a recombinase.

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Section: F a García-vázquez And Otherssupporting

confidence: 93%

Production of transgenic piglets using ICSI–sperm-mediated gene transfer in combination with recombinase RecA

García–Vázquez¹,

Ruiz²,

Matás³

et al. 2010

Reproduction

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“…Interestingly, the uptake of the DNA, at least in mammals, does not generally interfere with physiological semen parameters, such as motility at time of collection and progressive motility. In some cases, treated sperm cells perform better than untreated cells (Chan 2000).…”

Section: Technical Aspects Of the Sperm-mediated Gene Transfer Methodsmentioning

confidence: 99%

Sperm-mediated gene transfer

Chan¹,

Cm²,

Schatten³

2000

Current Topics in Developmental Biology

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Sperm-Mediated Gene Transfer

Lavitrano

Lulli²,

Maione³

et al. 1998

Microinjection and Transgenesis

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Since 1989, a new method for the production of transgenic animals has been available, namely spermmediated gene transfer (SMGT), based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA molecules and to transfer them into the oocyte at fertilisation. We first described the SMGT procedure in a small animal model, with high efficiency reported in the mouse. In addition, we successfully adapted and optimised the technique for use in large animals; it was, in fact, highly efficient in the generation of human decay accelerating factor transgenic pig lines, as well as multigene transgenic pigs in which three different reporter genes, namely enhanced green fluorescent protein, enhanced blue fluorescent protein and red fluorescent protein, were introduced. The major benefits of the SMGT technique were found to be its high efficiency, low cost and ease of use compared with other methods. Furthermore, SMGT does not require embryo handling or expensive equipment. Sperm-mediated gene transfer could also be used to generate multigene transgenic pigs that would be of benefit as large animal models for medical research, for agricultural and pharmaceutical applications and, in particular, for xenotransplantation, which requires extensive genetic manipulation of donor pigs to make them suitable for grafting to humans.

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Sperm-mediated DNA transfer to cells of the uterus and embryo

Cited by 7 publications

References 40 publications

Production of transgenic piglets using ICSI–sperm-mediated gene transfer in combination with recombinase RecA

Production of transgenic piglets using ICSI–sperm-mediated gene transfer in combination with recombinase RecA

Sperm-mediated gene transfer

Sperm-Mediated Gene Transfer

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