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 | |
| State | Published - Jan 4 2013 |
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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 journal › Article
}
TY - JOUR
T1 - Knockdown of recA gene expression by artificial small RNAs in Escherichia coli
AU - Sharma, Vandana
AU - Sakai, Yuta
AU - Smythe, Kiah A.
AU - Yokobayashi, Yohei
PY - 2013/1/4
Y1 - 2013/1/4
N2 - 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.
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
KW - Artificial small RNA
KW - Noncoding RNA
KW - RecA
KW - Swarming motility
UR - http://www.scopus.com/inward/record.url?scp=84872401866&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84872401866&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2012.10.141
DO - 10.1016/j.bbrc.2012.10.141
M3 - Article
VL - 430
SP - 256
EP - 259
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -