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

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

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.

MeSH terms

  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / genetics*
  • Escherichia coli Proteins / metabolism
  • Gene Expression Regulation, Bacterial*
  • Gene Library
  • Gene Silencing*
  • Genes, Reporter
  • Genetic Vectors / genetics
  • RNA, Small Untranslated / genetics*
  • RNA, Small Untranslated / metabolism

Substances

  • Escherichia coli Proteins
  • RNA, Small Untranslated