title: Artificial trans-encoded small non-coding RNAs specifically silence the selected gene expression in bacteria.
abstract: Recently, many small non-coding RNAs (sRNAs) with important regulatory roles have been identified in bacteria. As their eukaryotic counterparts, a major class of bacterial trans-encoded sRNAs acts by basepairing with target mRNAs, resulting in changes in translation and stability of the mRNA. RNA interference (RNAi) has become a powerful gene silencing tool in eukaryotes. However, such an effective RNA silencing tool remains to be developed for prokaryotes. In this study, we described first the use of artificial trans-encoded sRNAs (atsRNAs) for specific gene silencing in bacteria. Based on the common structural characteristics of natural sRNAs in Gram-negative bacteria, we developed the designing principle of atsRNA. Most of the atsRNAs effectively suppressed the expression of exogenous EGFP gene and endogenous uidA gene in Escherichia coli. Further studies demonstrated that the mRNA base pairing region and AU rich Hfq binding site were crucial for the activity of atsRNA. The atsRNA-mediated gene silencing was Hfq dependent. The atsRNAs led to gene silencing and RNase E dependent degradation of target mRNA. We also designed a series of atsRNAs which targeted the toxic genes in Staphyloccocus aureus, but found no significant interfering effect. We established an effective method for specific gene silencing in Gram-negative bacteria.
authors.affiliation: Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Man
authors.forename: Rubin
authors.initials: R
authors.lastname: Cheng
authors.forename: Cuicui
authors.initials: C
authors.lastname: Miao
authors.forename: Qianhong
authors.initials: Q
authors.lastname: Gong
authors.forename: Yuchao
authors.initials: Y
authors.lastname: Gu
authors.forename: Xinzhi
authors.initials: X
authors.lastname: Lu
authors.forename: Feng
authors.initials: F
authors.lastname: Han
authors.forename: Wengong
authors.initials: W
authors.lastname: Yu
mesh_terms: D020029:Base Pairing; D001665:Binding Sites; D004722:Endoribonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D005798:Genes, Bacterial; D020043:Genes, Essential; D005818:Genetic Engineering / Q000379:methods; D035001:Host Factor 1 Protein / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000737:chemistry; D058727:RNA, Small Untranslated / Q000737:chemistry*
chemical_list: D029968:Escherichia coli Proteins; C087373:Hfq protein, E coli; D035001:Host Factor 1 Protein; D012333:RNA, Messenger; D058727:RNA, Small Untranslated; D004722:Endoribonucleases; C022537:ribonuclease E