Subtracting Gene Function by Gene Silencing and Disruption in Bacteria

Nobutaka      Nakashima; Shan      Goh; Tomohiro      Tamura

Abstract

Gene silencing using antisense RNA or RNA interference (RNAi) is now a popular method used in eukaryotes. However, RNAi mechanism is absent in bacteria, and hence, single-stranded antisense RNAs, DNAs or nucleic acid analogs are used. Gene silencing in bacteria are achieved by either one of the following two approaches: expressing antisense RNAs using expression plasmids or adding synthetic antisense agents into the culture media. Recently, many advances on this method have been reported. In particular, the problem of low silencing efficiency has been solved remarkably, and as a result, this method is being used practically in biotechnology. This review mainly deals with the features and applications of gene silencing as a gene function subtracting method. Also, gene disruption is described as a competing and mutually complementary method.

Keywords: Antisense RNA, Escherichia coli, gene disruption, gene knock-down, gene knock-out, gene silencing, growth essential gene, LNA, locked nucleic acid, paired-termini, peptide nucleic acid, PNA, RNA silencing, ribosome-binding site.