Efficient and simple production of transgenic mice and rabbits using the new DMSO-sperm mediated exogenous DNA transfer method

A high efficient and simple transgenic technology on mice and rabbits to transfect spermatozoa with exogenous DNA/DMSO complex to obtain transgenic offspring, which is namely called DMSO‐sperm mediated gene transfer (SMGT). Mouse sperm could be either directly transfected via injection into testis o...

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Bibliographic Details
Published in:Molecular reproduction and development 2006-05, Vol.73 (5), p.589-594
Main Authors: Shen, Wei, Li, Lan, Pan, Qingjie, Min, Lingjiang, Dong, Huansheng, Deng, Jixian
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biotechnology
Cells, Cultured
Dimethyl Sulfoxide - chemistry
DMSO
Female
Fundamental and applied biological sciences. Psychology
Gene Transfer Techniques
Genetic engineering
Genetic technics
In Vitro Techniques
liposome
Male
Methods. Procedures. Technologies
Mice
Mice, Inbred ICR
Mice, Transgenic
mouse
Plasmids - chemistry
rabbit
Rabbits
spermatozoa
Spermatozoa - cytology
transgenesis
Transgenic animals
Transgenic animals and transgenic plants
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Summary:A high efficient and simple transgenic technology on mice and rabbits to transfect spermatozoa with exogenous DNA/DMSO complex to obtain transgenic offspring, which is namely called DMSO‐sperm mediated gene transfer (SMGT). Mouse sperm could be either directly transfected via injection into testis or cultured in vitro with the plasmed DNA containing the enhanced green fluorescent protein (EGFP) that could be expressed in the embryos and offspring. Then, 36 living transgenic rabbits were produced using the same technology, and the transgenic ratio of 56.3% was detected using PCR and Southern blot. As the controls, the transgenic ratios of 39.6% and 47.8% have also been tested using the liposomes mediated technology of Tfx™‐50 Reagent or Lipefectamin™‐2000, respectively. The results show that the female transgenic rabbits, as the mammary gland bioreactor models, could express the human tissue plasminogen activator mutant (htPAm) in their mammary cells when they are adult. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
ISSN:1040-452X
1098-2795
DOI:10.1002/mrd.20401