Directed Screening for sRNA Targets in E. coli Using a Plasmid Library

A large number of bacterial small regulatory RNAs (sRNAs) modulate gene expression by base pairing to a target mRNA, affecting its translation or stability. This posttranscriptional regulation has been shown to be essential and critical for bacterial physiology. One of the challenges of studying sRNA signaling is identifying the sRNA regulators of specific genes. Here, we describe a protocol for making an sRNA expression library and using this library to screen for sRNA regulators of genes of interest in E. coli. This library can be easily expanded and adapted to use in other bacteria.