Laboratório de Embriologia Molecular e Transgênese, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Theriogenology. 2011 May;75(8):1476-81. doi: 10.1016/j.theriogenology.2011.01.009. Epub 2011 Mar 9.
The objective was to introduce exogenous DNA into commercially sex-sorted bovine sperm using nanopolymer for transfection. In the first experiment, the optimal concentration and ratio of linear-to-circular plasmid was determined for NanoSMGT in unsorted sperm. A second experiment was conducted to transfect exogenous DNA into sex-sorted sperm. Exogenous DNA uptake occurred in a dose-dependent manner (P < 0.05). The optimal amount of DNA was 10 μg/10(6) cells. The ratios of linear-to-circular plasmid do not influence the uptake by unsorted sperm cells and none of the tested treatments affected sperm motility and viability. Commercially sex-sorted bovine sperm were able to uptake exogenous DNA using nanopolymer; however, both X- and Y-sorted sperm had decreased DNA uptake in comparison to unsorted sperm (P < 0.05). Neither sperm motility nor viability were affected by nanotransfection. In conclusion, nanopolymer efficiently introduced exogenous DNA into commercially sex-sorted bovine sperm; we inferred that these sperm could be used for production of embryos of the desired sex, a technique named NanoSMGT.
目的是使用纳米聚合物将外源 DNA 转染到商业性别分选的牛精子中。在第一个实验中，确定了线性到环形质粒的最佳浓度和比例用于未分选精子中的 NanoSMGT。第二个实验是将外源 DNA 转染到性别分选的精子中。外源性 DNA 的摄取呈剂量依赖性（P<0.05）。最佳 DNA 量为 10μg/10(6)细胞。线性到环形质粒的比例不会影响未分选精子的摄取，并且未测试的处理均未影响精子的活力和活力。商业性别分选的牛精子能够使用纳米聚合物摄取外源 DNA；然而，与未分选的精子相比，X 和 Y 分选的精子的 DNA 摄取量均减少（P<0.05）。纳米转染均未影响精子活力和活力。总之，纳米聚合物有效地将外源 DNA 导入商业性别分选的牛精子中；我们推断这些精子可用于生产所需性别的胚胎，该技术称为 NanoSMGT。
