ArticleNucleic acids research2009
Conditional gene silencing of multiple genes with antisense RNAs and generation of a mutator strain of Escherichia coli.
Article in Nucleic acids research, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed, 91 citations in OpenAlex.
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- Effect of Target Gene Silencing on Calcite Single Crystal Formation by Thermophilic Bacterium Geobacillus thermoglucosidasius NY05.Current microbiology · 2019Article
- Advances in engineered trans-acting regulatory RNAs and their application in bacterial genome engineering.Journal of industrial microbiology & biotechnology · 2019Review
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- Exploring of the feature space of de novo developed post-transcriptional riboregulators.PLoS computational biology · 2018Article
- The curli regulator CsgD mediates stationary phase counter-silencing of csgBA in Salmonella Typhimurium.Molecular microbiology · 2018Article
- Regulatory sRNAs in Cyanobacteria.Frontiers in microbiology · 2018Review
- Re-direction of carbon flux to key precursor malonyl-CoA via artificial small RNAs in photosyntheticBiotechnology for biofuels · 2018Article
- Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF.MicrobiologyOpen · 2017Article
- Efficient de novo synthesis of resveratrol by metabolically engineered Escherichia coli.Journal of industrial microbiology & biotechnology · 2017Article
- Optimization of a novel biophysical model using large scale in vivo antisense hybridization data displays improved prediction capabilities of structurally accessible RNA regions.Nucleic acids research · 2017Article
- Rational modular design of metabolic network for efficient production of plant polyphenol pinosylvin.Scientific reports · 2017Article
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Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In this study, we describe a method of simultaneous conditional gene silencing of up to four genes in Escherichia coli by using antisense RNAs. We used antisense RNAs with paired termini, which carried flanking inverted repeats to create paired double-stranded RNA termini; these RNAs have been proven to have high silencing efficacy. To express antisense RNAs, we constructed four IPTG-inducible vectors carrying different but compatible replication origins. When the lacZ antisense RNA was expressed using these vectors, lacZ expression was successfully silenced by all the vectors, but the expression level of the antisense RNA and silencing efficacy differed depending on the used vectors. All the vectors were co-transformable; the antisense RNAs against lacZ, ackA, pta and pepN were co-expressed, and silencing of all the target genes was confirmed. Furthermore, when antisense RNAs were targeted to the mutator genes mutS, mutD (dnaQ) and ndk, which are involved in DNA replication or DNA mismatch repair, spontaneous mutation frequencies increased over 2000-fold. The resulting mutator strain is useful for random mutagenesis of plasmids. The method provides a robust tool for investigating functional relationships between multiple genes or altering cell phenotypes for biotechnological and industrial applications.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.