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

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

Research output: Contribution to journalArticle

18 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)256-259
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume430
Issue number1
DOIs
StatePublished - Jan 4 2013

Fingerprint

Gene expression
Escherichia coli
RNA
Gene Expression
Bacterial RNA
Anti-Bacterial Agents
Small Untranslated RNA
DNA
Ciprofloxacin
DNA Repair
DNA Damage
Libraries
Repair
Cells
Phenotype
Messenger RNA
Pharmaceutical Preparations

Keywords

  • Antibiotic sensitivity
  • Artificial small RNA
  • Noncoding RNA
  • RecA
  • Swarming motility

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Knockdown of recA gene expression by artificial small RNAs in Escherichia coli. / Sharma, Vandana; Sakai, Yuta; Smythe, Kiah A.; Yokobayashi, Yohei.

In: Biochemical and Biophysical Research Communications, Vol. 430, No. 1, 04.01.2013, p. 256-259.

Research output: Contribution to journalArticle

Sharma, Vandana ; Sakai, Yuta ; Smythe, Kiah A. ; Yokobayashi, Yohei. / Knockdown of recA gene expression by artificial small RNAs in Escherichia coli. In: Biochemical and Biophysical Research Communications. 2013 ; Vol. 430, No. 1. pp. 256-259.
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