Team:Uppsala University/Translational
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IntaRNA, an RNA-RNA-interaction prediction software adapted for sRNA and ncRNA interactions [1] was used to predict the sRNA-mRNA interactions of the candidate sRNAs against the target mRNA containing the fusion between AAC(6’)-5’UTR. See <a href="https://2012.igem.org/Team:Uppsala_University/Modelling">modelling page</a> for details. Some of the sRNAs corresponding to the highest SYFP2 downregulation showed a significant basepair matching close to the RBS of the AAC(6’)-5’UTR. A few of the sRNAs was predicted to hybridize in the SYFP2 region of the mRNA.</p><br><br> | IntaRNA, an RNA-RNA-interaction prediction software adapted for sRNA and ncRNA interactions [1] was used to predict the sRNA-mRNA interactions of the candidate sRNAs against the target mRNA containing the fusion between AAC(6’)-5’UTR. See <a href="https://2012.igem.org/Team:Uppsala_University/Modelling">modelling page</a> for details. Some of the sRNAs corresponding to the highest SYFP2 downregulation showed a significant basepair matching close to the RBS of the AAC(6’)-5’UTR. A few of the sRNAs was predicted to hybridize in the SYFP2 region of the mRNA.</p><br><br> | ||
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Antibiotic resistance downregulation | Antibiotic resistance downregulation | ||
The next proof of concept was to test if the isolated sRNA also showed downregulation of the actual antibiotic resistance gene. The graphs below from the Etests [] show that the antibiotic resistance gene was repressed. This supports the hypothesis that it is the actual 5’UTR that is the key to control the the expression of the gene with sRNA.</p><br> | The next proof of concept was to test if the isolated sRNA also showed downregulation of the actual antibiotic resistance gene. The graphs below from the Etests [] show that the antibiotic resistance gene was repressed. This supports the hypothesis that it is the actual 5’UTR that is the key to control the the expression of the gene with sRNA.</p><br> | ||
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+ | <b>sRNA UU17</b> | ||
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+ | <a href="https://static.igem.org/mediawiki/2012/7/7d/SRNA17-mRNA_medium.png"><img src="https://static.igem.org/mediawiki/2012/7/7d/SRNA17-mRNA_medium.png" id="intpic"></a> | ||
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Revision as of 02:31, 27 September 2012
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References |
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[1] Sharma, V., Yamamura, A., Yokobayashi, Y., 2012. Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli. ACS Synth. Biol. 1, 6–13.
[2] Holmqvist, E., Unoson, C., Reimegård, J., Wagner, E.G.H., 2012. A mixed double negative feedback loop between the sRNA MicF and the global regulator Lrp. Molecular Microbiology 84, 414–427.