由tRNA基因组成的障碍物可以补充无处不在的染色体开放元件的好处,以增强转基因表达
Rebecca E Sizer1, Richard M Ingram1, Robert J White1
1Department of Biology, University of York, York, UK.
Biotechnology journal
|February 16, 2025
概括
我们发现,与无处不在的染色体开放元素 (UCOEs) 相比,tRNA基因在生物制造中提供了对转基因沉默的优越屏障. 结合两者的混合元素可能会增强长期的基因表达.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 转基因整合到不稳定的基因组区域导致生物制造中不可靠的表达.
- 屏障元素用于改善转基因的稳定性和表达.
- 无处不在的染色体开放元件 (UCOE) 是一个常见的屏障元件.
研究的目的:
- 为了比较tRNA基因对UCOE在中国仓鼠卵巢 (CHO) 细胞中的屏障疗效.
- 调查基因表达稳定性背后的表观遗传机制.
- 探索混合屏障元件在生物制造中的潜力.
主要方法:
- 屏障元素 (UCOE,tRNA基因) 与CHO细胞中的记者转基因的整合.
- 长期细胞培养和对转基因表达水平的分析.
- 染色体免疫沉 (ChIP) 来评估表观遗传标记 (H3K9me3,H3K27ac) 和RNA聚合酶II的招募.
主要成果:
- 在CHO细胞中,tRNA基因比UCOE对转基因沉默提供了更有效的屏障.
- UCOE强烈增强了表达力,但是暂时的,并且减少了变性.
- 这两种元素都抵消了H3K9me3;只有tRNA基因维持了H3K27ac和RNA聚合酶II的长期招募.
结论:
- 与UCOE相比,tRNA基因提供了优越的长期转基因沉默保护.
- 结合UCOE和tRNA基因益处的合成混合元件可以增强生物制造的持久转基因表达.
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