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IGEM:Caltech/2007/Project/Riboregulator: Difference between revisions
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Revision as of 05:25, 26 October 2007
, 26 October 2007→Riboregulator Design
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Our riboregulators consist of two interacting parts: cis and trans. The cis sequence is located downstream from the promoter to a gene of interest and upstream of the ribosome binding site (RBS). This cis sequence is complementary to the RBS and forms a stem-loop at the 5’ end of the mRNA after transcription, thus blocking ribosome binding and translation. Gene expression is turned on with a trans noncoding RNA. This trans sequence is produced from another promoter in a different plasmid and targets the cis-repressed RNA with high specificity. When the trans transcript is available, it binds to the cis region and thus opens the RBS for ribosome docking and subsequent translation. | Our riboregulators consist of two interacting parts: cis and trans. The cis sequence is located downstream from the promoter to a gene of interest and upstream of the ribosome binding site (RBS). This cis sequence is complementary to the RBS and forms a stem-loop at the 5’ end of the mRNA after transcription, thus blocking ribosome binding and translation. Gene expression is turned on with a trans noncoding RNA. This trans sequence is produced from another promoter in a different plasmid and targets the cis-repressed RNA with high specificity. When the trans transcript is available, it binds to the cis region and thus opens the RBS for ribosome docking and subsequent translation. | ||
The cis repressive sequences are approximately 50 base pairs and do not alter the reading frame of the native gene. The trans RNA is approximately 90 base pairs and contains a nucleotide sequence that is complementary to the cis RNA. RNAstructure3 software was used to predict the secondary structure as well as the free energy of the cis and trans elements and their interactions. | The cis repressive sequences are approximately 50 base pairs and do not alter the reading frame of the native gene. The trans RNA is approximately 90 base pairs and contains a nucleotide sequence that is complementary to the cis RNA. RNAstructure3 software was used to predict the secondary structure as well as the free energy of the cis and trans elements and their interactions. | ||
[[Image:Cis trans diagram.JPG|left|thumb|500px|]] | |||
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[[Image:Trans designs 1.JPG|left|thumb| | [[Image:Trans designs 1.JPG|left|thumb|600px|The 5 trans-activating riboregulators are shown with purple bases indicating regions of complimentarity to the cis-repressive riboregulators.]] | ||
[[Image:Cis trans compliments.JPG|right|thumb|500px|A schematic diagram showing the parts of cis1 repressor and trans1 activator that are complimentary to each other. Note that the RBS is now open.]] | |||
==Experiments with Riboregulators== | ==Experiments with Riboregulators== | ||
