• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Gene expression >Gene expression pattern and downregulation of DNA methyltransferase 1 using siRNA in porcine somatic cells.
【24h】

Gene expression pattern and downregulation of DNA methyltransferase 1 using siRNA in porcine somatic cells.

机译：猪体细胞中使用siRNA的基因表达模式和DNA甲基转移酶1的下调。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

DNA methylation plays a significant role in the expression of the genetic code and affects early growth and development through their influence on gene expression. Manipulation of the DNA methylation marks of differentiated cells will allow a better understanding of the different molecular processes associated with chromatin structure and gene expression. The objective of this study was to identify small interfering RNAs (siRNAs) with the ability to reduce DNA methyltransferase 1 (Dnmt1) mRNA and consequently decrease Dnmt1 protein as well as DNA methylation in porcine cells. Fibroblasts from four porcine fetuses were established and cultured in 5% CO2 in air at 38 degrees C. Optimal transfection conditions were evaluated using a FITC-labeled control siRNA. Four Dnmt1-specific siRNAs were evaluated upon transfection of each cell line. A nonsilencing siRNA was used as a negative control. The expression patterns of Dnmt1 were analyzed by Q-PCR. The combination of 1 microg of siRNA and a 1:6 siRNA to transfection reagent ratio produced the highest transient transfection rates without affecting cell viability. Downregulation of Dnmt1 varied between siRNAs. Transfection of porcine cells with highly effective siRNAs resulted in a drastic reduction of Dnmt1 mRNA and a slight decrease in protein production. However, this small reduction in the protein concentration induced significant genomic hypomethylation. These data suggest that although Dnmt1 mRNA abundance plays an important role during protein regulation, Dnmt1 enzyme is mainly posttranscriptionally regulated. Subsequent use of these cells for cloning, differentiation, and cancer studies will provide insight as to how methylation of the DNA affects genomic reprogramming.
机译：DNA甲基化在遗传密码的表达中起着重要作用，并通过影响基因表达而影响早期的生长发育。操纵分化细胞的DNA甲基化标记可以更好地理解与染色质结构和基因表达有关的不同分子过程。这项研究的目的是鉴定具有减少DNA甲基转移酶1（Dnmt1）mRNA从而减少Dnmt1蛋白以及猪细胞DNA甲基化的能力的小干扰RNA（siRNA）。建立了来自四个猪胎儿的成纤维细胞，并在38°C的空气中于5％CO2中培养。使用FITC标记的对照siRNA评估了最佳转染条件。每种细胞系转染后评估了四个Dnmt1特异性siRNA。非沉默的siRNA用作阴性对照。通过Q-PCR分析Dnmt1的表达模式。 1微克siRNA和1：6 siRNA与转染试剂的比例组合产生了最高的瞬时转染速率，而不会影响细胞活力。 Dnmt1的下调在siRNA之间有所不同。用高效siRNA转染猪细胞导致Dnmt1 mRNA急剧减少，蛋白质产量略有下降。但是，蛋白质浓度的这种小幅下降会引起显着的基因组低甲基化。这些数据表明，尽管Dnmt1 mRNA的丰度在蛋白质调节过程中起着重要作用，但Dnmt1酶主要是转录后调节的。随后将这些细胞用于克隆，分化和癌症研究将提供有关DNA甲基化如何影响基因组重编程的见解。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号