工程化传感器用于基因表达负担的检测。

Engineering Sensors for Gene Expression Burden.

机构信息

Department of Bioengineering, Imperial College London, London, UK.

Imperial College Centre for Synthetic Biology, Imperial College London, London, UK.

出版信息

Methods Mol Biol. 2021;2229:313-330. doi: 10.1007/978-1-0716-1032-9_15.

DOI:10.1007/978-1-0716-1032-9_15

PMID:33405229

Abstract

RNA-seq enables the analysis of gene expression profiles across different conditions and organisms. Gene expression burden slows down growth, which results in poor predictability of gene constructs and product yields. Here, we describe how we applied RNA-seq to study the transcriptional profiles of Escherichia coli when burden is elicited during heterologous gene expression. We then present how we selected early responsive promoters from our RNA-seq results to design sensors for gene expression burden. Finally, we describe how we used one of these sensors to develop a burden-driven feedback regulator to improve cellular fitness in engineered E. coli.

摘要

RNA-seq 能够分析不同条件和生物体的基因表达谱。基因表达负担会减缓生长速度，从而导致基因构建体和产物产量的可预测性较差。在这里，我们描述了如何在异源基因表达过程中引发基因负担时，应用 RNA-seq 来研究大肠杆菌的转录谱。然后，我们介绍了如何从 RNA-seq 结果中选择早期响应启动子来设计用于基因表达负担的传感器。最后，我们描述了如何使用其中一个传感器来开发负担驱动的反馈调节剂，以改善工程大肠杆菌中的细胞适应性。