Inducible Expression of dsRNA in Escherichia coli

Methods Mol Biol. 2024:2771:57-64. doi: 10.1007/978-1-0716-3702-9_9.

Abstract

Double-stranded RNA (dsRNA) is a valuable tool for reverse genetics research and gene silencing applications. It is also an important management method for pests and diseases in agriculture. It can be synthesized both in vivo and in vitro. The latter presents the drawback of high production cost, the former is less expensive and suitable for scalable production. In general, dsRNAs are obtained in vivo from Escherichia coli heterologous systems that require the IPTG-inducible T7 RNA polymerase. In this report, we describe the construction of an RNAi system for the expression of dsRNA using the HT115 bacterial strain and the L4440 plasmid, and the extraction and identification of dsRNA.

Keywords: Escherichia coli; In vivo synthesis; dsRNA synthesis.

MeSH terms

  • Agriculture*
  • Escherichia coli / genetics
  • Gene Silencing
  • RNA Interference
  • RNA, Double-Stranded* / genetics

Substances

  • RNA, Double-Stranded