Knockdown of recA gene expression by artificial small RNAs in Escherichia coli

Vandana Sharma; Yuta Sakai; Kiah A. Smythe; Yohei Yokobayashi

doi:10.1016/j.bbrc.2012.10.141

Knockdown of recA gene expression by artificial small RNAs in Escherichia coli

Vandana Sharma, Yuta Sakai, Kiah A. Smythe, Yohei Yokobayashi

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

Bacterial RecA plays a central role in DNA repair and regulation of the SOS response to DNA damage, and has been suggested as a new antibiotic drug target. To develop a new tool to study RecA function, we engineered artificial small RNAs (sRNAs) that can posttranscriptionally repress RecA expression in Escherichia coli. The artificial sRNAs mimic the bacterial noncoding sRNAs which possess an antisense domain that is partially complementary to the targeted mRNA. We screened a library of artificial sRNAs with a randomized antisense domain and isolated several anti-recA sRNAs that can knockdown the endogenous RecA level in E. coli. The cells expressing the anti-recA sRNAs were found to exhibit phenotypes consistent with RecA repression such as reduced swarming motility and increased susceptibility to ciprofloxacin, a fluoroquinone antibiotic.

Original language	English (US)
Pages (from-to)	256-259
Number of pages	4
Journal	Biochemical and Biophysical Research Communications
Volume	430
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2012.10.141
State	Published - Jan 4 2013

Keywords

Antibiotic sensitivity
Artificial small RNA
Noncoding RNA
RecA
Swarming motility

ASJC Scopus subject areas

Biochemistry
Biophysics
Cell Biology
Molecular Biology

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abstract = "Bacterial RecA plays a central role in DNA repair and regulation of the SOS response to DNA damage, and has been suggested as a new antibiotic drug target. To develop a new tool to study RecA function, we engineered artificial small RNAs (sRNAs) that can posttranscriptionally repress RecA expression in Escherichia coli. The artificial sRNAs mimic the bacterial noncoding sRNAs which possess an antisense domain that is partially complementary to the targeted mRNA. We screened a library of artificial sRNAs with a randomized antisense domain and isolated several anti-recA sRNAs that can knockdown the endogenous RecA level in E. coli. The cells expressing the anti-recA sRNAs were found to exhibit phenotypes consistent with RecA repression such as reduced swarming motility and increased susceptibility to ciprofloxacin, a fluoroquinone antibiotic.",

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AB - Bacterial RecA plays a central role in DNA repair and regulation of the SOS response to DNA damage, and has been suggested as a new antibiotic drug target. To develop a new tool to study RecA function, we engineered artificial small RNAs (sRNAs) that can posttranscriptionally repress RecA expression in Escherichia coli. The artificial sRNAs mimic the bacterial noncoding sRNAs which possess an antisense domain that is partially complementary to the targeted mRNA. We screened a library of artificial sRNAs with a randomized antisense domain and isolated several anti-recA sRNAs that can knockdown the endogenous RecA level in E. coli. The cells expressing the anti-recA sRNAs were found to exhibit phenotypes consistent with RecA repression such as reduced swarming motility and increased susceptibility to ciprofloxacin, a fluoroquinone antibiotic.

KW - Antibiotic sensitivity

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KW - Noncoding RNA

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KW - Swarming motility

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Knockdown of recA gene expression by artificial small RNAs in Escherichia coli

Abstract

Keywords

ASJC Scopus subject areas

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