Abstract
It has become increasingly evident that noncoding small RNAs (sRNAs) play a significant and global role in bacterial gene regulation. A majority of the trans-acting sRNAs in bacteria interact with the 50 untranslated region (UTR) and/ or the translation initiation region of the targeted mRNAs via imperfect base pairing, resulting in reduced translation efficiency and/or mRNA stability. Additionally, bacterial sRNAs often contain distinct scaffolds that recruit RNA chaperones such as Hfq to facilitate gene regulation. In this study, we describe a strategy to engineer artificial sRNAs that can regulate desired endogenous genes in Escherichia coli. Using a fluorescent reporter gene that was translationally fused to a native 50 mRNA leader sequence, active artificial sRNAs were screened from libraries in which natural sRNA scaffolds were fused to a randomized antisense domain. Artificial sRNAs that posttranscriptionally repress two endogenous genes ompF and fliC were isolated and characterized. We anticipate that the artificial sRNAs will be useful for dynamic control and fine-tuning of endogenous gene expression in bacteria for applications in synthetic biology.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6-13 |
| Number of pages | 8 |
| Journal | ACS Synthetic Biology |
| Volume | 1 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 20 2012 |
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Keywords
- Gene knockdown
- Gene regulation
- Noncoding RNA
- RNA engineering
- Small RNA
- Translational regulation
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
- Biomedical Engineering
Cite this
Engineering artificial small RNAs for Conditional gene silencing in escherichia coli. / Sharma, Vandana; Yamamura, Asami; Yokobayashi, Yohei.
In: ACS Synthetic Biology, Vol. 1, No. 1, 20.01.2012, p. 6-13.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Engineering artificial small RNAs for Conditional gene silencing in escherichia coli
AU - Sharma, Vandana
AU - Yamamura, Asami
AU - Yokobayashi, Yohei
PY - 2012/1/20
Y1 - 2012/1/20
N2 - It has become increasingly evident that noncoding small RNAs (sRNAs) play a significant and global role in bacterial gene regulation. A majority of the trans-acting sRNAs in bacteria interact with the 50 untranslated region (UTR) and/ or the translation initiation region of the targeted mRNAs via imperfect base pairing, resulting in reduced translation efficiency and/or mRNA stability. Additionally, bacterial sRNAs often contain distinct scaffolds that recruit RNA chaperones such as Hfq to facilitate gene regulation. In this study, we describe a strategy to engineer artificial sRNAs that can regulate desired endogenous genes in Escherichia coli. Using a fluorescent reporter gene that was translationally fused to a native 50 mRNA leader sequence, active artificial sRNAs were screened from libraries in which natural sRNA scaffolds were fused to a randomized antisense domain. Artificial sRNAs that posttranscriptionally repress two endogenous genes ompF and fliC were isolated and characterized. We anticipate that the artificial sRNAs will be useful for dynamic control and fine-tuning of endogenous gene expression in bacteria for applications in synthetic biology.
AB - It has become increasingly evident that noncoding small RNAs (sRNAs) play a significant and global role in bacterial gene regulation. A majority of the trans-acting sRNAs in bacteria interact with the 50 untranslated region (UTR) and/ or the translation initiation region of the targeted mRNAs via imperfect base pairing, resulting in reduced translation efficiency and/or mRNA stability. Additionally, bacterial sRNAs often contain distinct scaffolds that recruit RNA chaperones such as Hfq to facilitate gene regulation. In this study, we describe a strategy to engineer artificial sRNAs that can regulate desired endogenous genes in Escherichia coli. Using a fluorescent reporter gene that was translationally fused to a native 50 mRNA leader sequence, active artificial sRNAs were screened from libraries in which natural sRNA scaffolds were fused to a randomized antisense domain. Artificial sRNAs that posttranscriptionally repress two endogenous genes ompF and fliC were isolated and characterized. We anticipate that the artificial sRNAs will be useful for dynamic control and fine-tuning of endogenous gene expression in bacteria for applications in synthetic biology.
KW - Gene knockdown
KW - Gene regulation
KW - Noncoding RNA
KW - RNA engineering
KW - Small RNA
KW - Translational regulation
UR - http://www.scopus.com/inward/record.url?scp=84867191934&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84867191934&partnerID=8YFLogxK
U2 - 10.1021/sb200001q
DO - 10.1021/sb200001q
M3 - Article
C2 - 23651005
AN - SCOPUS:84867191934
VL - 1
SP - 6
EP - 13
JO - ACS Synthetic Biology
JF - ACS Synthetic Biology
SN - 2161-5063
IS - 1
ER -