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Title 

Exogenous DNA uptake of boar spermatozoa by a magnetic nanoparticle vector system

Authors 

T S KimS H LeeGil-Tae GangY S LeeS U KimD B KooM Y ShinC K ParkD S Lee

Publisher 

Wiley-Blackwell

Issue Date 

2010

Citation 

Reproduction in Domestic Animals, vol. 45, no. 5, pp. e201-e206

Abstract 

The sperm-mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore-labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.

ISSN 

0936-6768

Link 

http://dx.doi.org/10.1111/j.1439-0531.2009.01516.x

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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