In Vivo Screening of Artificial Small RNAs for Silencing Endogenous Genes in Escherichia coli

  • Vandana Sharma
  • Yohei Yokobayashi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1073)

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.

Key words

Noncoding RNA Small RNA Gene regulation Gene silencing RNA engineering Translational regulation 

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Vandana Sharma
    • 1
  • Yohei Yokobayashi
    • 1
  1. 1.Department of Biomedical EngineeringUniversity of CaliforniaDavisUSA

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