Testis-mediated gene transfer in mice: comparison of transfection reagents regarding transgene transmission and testicular damage
Biol. Res
;
44(3): 229-234, 2011. ilus, tab
Article
Dans Anglais
| LILACS
| ID: lil-608618
ABSTRACT
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. PC R 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.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Testicule
/
Souris transgéniques
/
Diméthylsulfoxyde
/
Techniques de transfert de gènes
/
Transgènes
/
Protéines à fluorescence verte
Limites du sujet:
Animaux
langue:
Anglais
Texte intégral:
Biol. Res
Thème du journal:
Biologie
Année:
2011
Type:
Article
/
descriptif de projet
Pays d'affiliation:
Brésil
Institution/Pays d'affiliation:
Unlversldade Federal de Pelotas/BR
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