Techniques for augmentation of exogenous DNA uptake by ovine spermatozoa

Authors

  • K. Hoseini Pajooh Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
  • M. Behdani Molecular Medicine Depart-ment, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • M. Karimipoor Molecular Medicine Depart-ment, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • P. Tajik Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
Abstract:

Sperm mediated gene transfer can be an inexpensive and simple method in animal transgenesis; however its efficiency is poor, mainly due to the spermatozoa’s lesser uptake of exogenous DNA. In the present study, the effects of lipofection and other augmentation techniques, such as sperm freezing and spermatozoa treatment with triton X100 and DMSO, on exogenous DNA uptake by sheep spermatozoa and motility of sperms with plasmid uptake were evaluated. In the first experiment, ram sperms were incubated with a complex of rhodamine labeled plasmid (p-EGFP) and Lipofectamine 2000TM. In the second, spermatozoa were treated with Triton X-100TM or DMSO or were frozen without cryoprotectant. The results indicated that there was no significant difference (P

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Journal title

volume 17  issue 1

pages  25- 30

publication date 2016-03-01

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