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Large‐loop antisense RNA hairpins provide stabilized and highly efficient antisense regulators in Escherichia coli
Author(s) -
Nakashima Nobutaka,
Tamura Tomohiro,
Good Liam
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a538-c
Subject(s) - antisense rna , escherichia coli , loop (graph theory) , rna , chemistry , microbiology and biotechnology , biology , genetics , gene , mathematics , combinatorics
Antisense RNAs (ASRs) transcribed from expression plasmids specifically hybridize to target mRNA and inhibit gene expression in bacteria. They are useful tools for functional genomics but in Escherichia coli , the ASR method has shown limited efficiency. We demonstrate that the inhibition efficiency can be improved greatly by placing ASR between an artificial inverted repeat (IR) sequence. In one example, the non‐essential ackA gene (encoding acetate kinase) was targeted and inhibited up to 75 %. RNA analysis showed that the ASR was abundant and stable and the target mRNA was degraded. In a second example, the essential fabI gene (encoding enol‐ACP reductase) was targeted and the antisense effect prevented growth and sensitize cells to triclosan. We also show some application examples of this new ASR method.