工程转录因子结合阵列用于基于DNA的哺乳动物细胞基因表达控制
A Zouein1,2,3, B Lende-Dorn4, K E Galloway4
1Department of Chemical Engineering, Imperial College London, London, UK.
bioRxiv : the preprint server for biology
|September 16, 2024
概括
研究人员开发了称为转录因子 (TF) 识别元素 (RE) 阵列的DNA工具,以精确控制哺乳动物细胞中的基因表达. 这些数组可以隔离TFs,为细胞工程和合成生物学应用提供了新的可能性.
科学领域:
- 合成生物学 合成生物学
- 分子和细胞生物学分子和细胞生物学.
- 基因工程是一种基因工程.
背景情况:
- 哺乳动物细胞工程依赖于精确控制基因表达.
- 转录因子 (TF) 是基因表达的关键调节者.
- 现有的TF操纵方法存在局限性.
研究的目的:
- 探索转录因子识别元素 (RE) 阵列作为调节哺乳动物细胞TF水平和基因表达的DNA工具.
- 开发一种用于组装长重复的RE数组的方法,用于TF封存.
- 为了证明RE阵列在控制合成和原生TF活动中的应用,用于细胞工程.
主要方法:
- 使用不同长度的Tet TF结合RE数组来调整基因表达的概念验证.
- 开发克隆麻烦的循环重复 (CTRL) 方法,以组装多达256次RE重复的等离子体.
- 将RE阵列等离子体转化为哺乳动物细胞,以评估TF分离和基因调节.
- 展示了针对合成和本地哺乳动物TF的RE阵列.
主要成果:
- Tet RE 阵列证明了基因表达和基因电路性能的可预测调整.
- 在CTRL方法成功地组装了大量RE重复的等离子体.
- 较长的RE数组大小显示了通过TF分离修改宿主细胞基因调节的潜力.
- 通过准TFs,RE阵列等离子体有效调节遗传电路并影响细胞命运.
结论:
- 结合TF的RE数组代表了一种用于操纵哺乳动物基因表达的新型DNA工具.
- CTRL 方法有助于构建复杂的重复DNA数组,用于生物应用.
- 这种方法扩大了哺乳动物细胞工程的工具包,使基因调节和细胞命运的确切控制成为可能.
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