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Team:Bologna/Project
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Introduction
Our goal is to create a logic gate based on a post-transcriptional regulation system in Escherichia coli, using RNA to silence gene expression. We inserted a cis-repressing sequence directly upstream of the rbs (ribosome binding site) in a target gene and we created the complementary trans-repressor sequence with 2 kind of rbs cover. We want to use the short non coding segment of RNA, that is expressed in trans, to cover the mRNA’s rbs. In this way the mRNA can't bind anymore with ribosomes, silencing gene expression. We developed a bioinformatic tool to research the sequences and these are the results:
| Cis-repressing sequence, inserted upstream of the target gene | |||
| Scar | Sequence | Rbs | Scar |
| TACTAGAG | AACACAAACTATCACTTTAACAACACATTACATATACATTAAAATATTAC | AAAGAGGAGAAA | TACTAGAG |
| Trans-repressor sequence with a cover of 7 bases (long version) | |||
| Scar | Cover | Trans(long) | Scar |
| TACTAGAG | CCTCTTT | GTAATATTTTAATGTATATGTAATGTGTTGTTAAAGTGATAGTTTGTGTT | TACTAGAG |
| Trans-repressor sequence with a cover of 4 bases (short version) (don’t cover Shine-Dalgarno box) | |||
| Scar | Cover | Trans(short) | Scar |
| TACTAGAG | TCTTT | GTAATATTTTAATGTATATGTAATGTGTTGTTAAAGTGATAGTTTGTGTT | TACTAGAG |
