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矩阵附着区域通过操纵稳定转染的CHO-K1细胞中的位置依赖效应来增强转基因表达

Jihong Zhang^1,2, Lin Wang¹, Xi Zhang²

¹School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453003, China.

Biochemistry and cell biology = Biochimie et biologie cellulaire

|July 19, 2024

概括

矩阵附着区域 (MAR) 的战略位置显著提高了重组蛋白质的生产. 带有MARs的侧边表达盒优化了CHO-K1细胞中的转基因表达和细胞生产率.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
基因工程是一种基因工程.

背景情况:

矩阵附着区域 (MAR) 是影响基因表达的关键DNA元素.
以前的研究表明,MAR位置会影响转基因表达,但缺乏系统的研究.

研究的目的:

系统地研究MARs对转基因表达的位置依赖性影响.
为了确定最佳的MAR放置,以增强重组蛋白质的生产.

主要方法:

构建带有MAR的向量 (MAR 1-68,MAR X-29) 在相对于促进子和表达卡塞特的不同位置.
传染CHO-K1细胞并量化增强绿色光蛋白 (eGFP) 表达水平和表达eGFP细胞的百分比.
在转染细胞中分析红色素 (EPO) 的生产力.
进行生物信息学分析以确定相关的转录因子.

主要成果:

位于促进体上游的MAR显著增加了EGFP表达水平.
在表达卡塞特旁边的MAR显示了eGFP表达的最实质性的增加 (97.8%和96.0%与没有MAR的73.7%相比).
搭配MAR的建筑物产生了最高的EPO生产率.

结论:

带有MARs围绕表达盒的载体是CHO-K1细胞中增强重组蛋白质生产的最佳选择.
战略性MAR在矢量设计中的整合对于优化重组蛋白表达至关重要.

关键词:

中国仓鼠的卵巢细胞.矩阵连接区域是矩阵连接区域.位置效应影响位置效应转基因表达的表达方式

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