在体内筛选用于沉默大肠杆菌内源性基因的人工小RNA。

In vivo screening of artificial small RNAs for silencing endogenous genes in Escherichia coli.

作者信息

Sharma Vandana, Yokobayashi Yohei

机构信息

Department of Biomedical Engineering, University of California, Davis, CA, USA.

出版信息

Methods Mol Biol. 2013;1073:75-84. doi: 10.1007/978-1-62703-625-2_8.

DOI:10.1007/978-1-62703-625-2_8

PMID:23996441

Abstract

Bacterial noncoding small RNAs (sRNAs) modulate expression of numerous genes through antisense interactions with mRNAs. This chapter describes an in vivo screening strategy to engineer artificial sRNAs that can posttranscriptionally regulate desired endogenous genes in Escherichia coli. Artificial sRNA libraries are constructed by randomizing the antisense domain of natural sRNAs and screened for gene silencing activity using a cotransformed reporter vector. These small synthetic riboregulators can be used in synthetic gene circuits to control cell functions by directly targeting endogenous genes.

摘要

细菌非编码小RNA（sRNA）通过与mRNA的反义相互作用调节众多基因的表达。本章描述了一种体内筛选策略，用于构建可在转录后调控大肠杆菌中所需内源基因的人工sRNA。通过随机化天然sRNA的反义结构域构建人工sRNA文库，并使用共转化的报告载体筛选基因沉默活性。这些小型合成核糖调节因子可用于合成基因电路，通过直接靶向内源基因来控制细胞功能。