title: An Effective Method for Specific Gene Silencing in Escherichia coli Using Artificial Small RNA.
abstract: Knockdown or silencing of a specific gene presents a powerful strategy for elucidating gene function in a variety of organisms. To date, efficient silencing methods have been established in eukaryotes, but not bacteria. In this chapter, an efficient and versatile gene silencing method using artificial small RNA (afsRNA) is described. For this purpose, target-recognizing sequences were introduced in specially designed RNA scaffolds to exist as single-stranded stretches in afsRNA. The translation initiation region of target genes was used as the sequence for afsRNA recognition, based on the theory that this site is usually highly accessible to ribosomes, and therefore, possibly, afsRNA. Two genes transcribed as monocistrons were tested with our protocol. Both genes were effectively silenced by their cognate afsRNAs.
authors.affiliation: Department of Chemistry, KAIST, Daejeon, 305-701, Korea.
authors.forename: Geunu
authors.initials: G
authors.lastname: Bak
authors.forename: Jee Soo
authors.initials: JS
authors.lastname: Choi
authors.forename: Wonkyeong
authors.initials: W
authors.lastname: Kim
authors.forename: Shinae
authors.initials: S
authors.lastname: Suk
authors.forename: Younghoon
authors.initials: Y
authors.lastname: Lee
mesh_terms: D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D009690:Nucleic Acid Conformation; D058727:RNA, Small Untranslated / Q000737:chemistry* / Q000235:genetics*
chemical_list: D058727:RNA, Small Untranslated