NanoSMGT: transfection of exogenous DNA on sex-sorted bovine sperm using nanopolymer

Campos, Vinícius Farias; Komninou, Eliza Rossi; Urtiaga, G.; Leon, P.M. de; Seixas, Fabiana Kömmling; Dellagostin, Odir A.; Deschamps, João Carlos; Collares, Tiago

doi:10.1016/j.theriogenology.2011.01.009

Cited by 34 publications

(17 citation statements)

References 34 publications

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“…The fi rst report that exogenous DNA could be introduced into sperm was made by Brackett et al (1971). Several studies in distinct species have reported the generation of transgenic animals using spermatozoa as vectors to carrier foreign DNA to the ova (Lu et al, 2002;Webster et al, 2005;Shen et al, 2006;Hoelker et al, 2007, Campos et al, 2011a, Campos et al, 2011b, Campos et al, 2011c. One approach of sperm-mediated gene transfer (SMGT) is the direct introduction of foreign DNA into testes, so-called testis-mediated gene transfer (TMGT), which allows for natural mating and mass gene transfer.…”

Section: Introductionmentioning

confidence: 99%

Testis-mediated gene transfer in mice: comparison of transfection reagents regarding transgene transmission and testicular damage

et al. 2011

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Testis-mediated gene transfer (TMGT) has been used as in vivo gene transfer technology to introduce foreign DNA directly into testes, allowing mass gene transfer to offspring via mating. In this study, we used plasmid DNA (pEGFP-N1) mixed with dimethylsulfoxide (DMSO), N,N-dimethylacetamide (DMA) or liposome (Lipofectin) in an attempt to improve TMGT. Males receiving consecutive DNA complex injections were mated to normal females to obtain F0 progeny. In vivo evaluation of EGFP expression, RT-PCR and PCR were used to detect the expression and the presence of exogenous DNA in the progeny. We also evaluated possible testicular damage by histological procedures. PCR and RT-PCR analyses revealed that liposome and DMSO increased the rate of TMGT. Histological analyses demonstrated that repeated (4 times) injections of DNA complexes can affect spermatogenesis. DMSO was the most deleterious among the reagents tested. In this study, we detected the presence of transgene in the progeny, and its expression in blood cells. Consecutive injections of DNA complexes were associated with impaired spermatogenesis, suggesting requirement of optimal conditions for DNA delivery through TMGT.

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Section: Introductionmentioning

confidence: 99%

Testis-mediated gene transfer in mice: comparison of transfection reagents regarding transgene transmission and testicular damage

et al. 2011

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“…Recently we have demonstrated the successful use of a cationic dendrimer for exogenous DNA transfection into sex-sorted bovine sperm [8]. In addition we demonstrated that this dendrimer was able to improve the number of internalized plasmids by sperm cells in comparison to other common transfection methods such as lipofection or incubation leading to a significant improvement in production of transgenic bovine embryos [7].…”

Section: Nanotubes For Dna Transfectionmentioning

confidence: 91%

“…Several approaches have been utilized in order to improve DNA uptake. Methods such as electroporation, lipofection, DNA/DMSO complexes and restriction-enzyme-mediated integration (REMI) have been used; however the frequency of transgenic offspring remains low [6,8,21,43]. Conversely, the more recent use of nanotechnology in the context of SMGT has new possibilities for the efficient delivery of exogenous DNA into sperm.…”

Section: Nanotubes For Dna Transfectionmentioning

confidence: 99%

The Use of Nanostructures for DNA Transfection

Campos

Yurgel

Seixas

et al. 2012

Carbon Nanostructures

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The interaction between nanostructured materials and living systems is of fundamental and practical interest and will determine the biocompatibility, potential utilities and applications of novel nanomaterials in biological settings. The pursuit of new types of molecular transporters is an active area of research, due to the high impermeability of cell membranes and other biological barriers to foreign substances and the need for intercellular delivery of molecules via cell-penetrating transporter for drug, gene or protein therapeutics. Here, is described the novel nanostructurebased transfection systems. The transfection uses of nanopolymers, nanoparticles and nanotubes are the main focus of this review. In addition are described the technique called NanoSMGT that uses nanostructures for DNA transfection in sperm cells that could be used for transgenic animal generation or human gene therapy.

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“…Actions of nanotransfectants, such as nanopolymers, are based on the sperm's inherent ability to spontaneously bind and incorporate exogenous DNA during co-incubation. Thereby they serve as natural vectors for transfection of the oocyte and, consequently, of a developing embryo via IVF (Campos et al, 2011b). This treatment did not affect sperm motility or viability, but both Xand Y-sorted sperm had decreased DNA uptake in comparison with unsorted sperm (Campos et al, 2011a).…”

Section: Embryogenesismentioning

confidence: 97%