Team:TUDelft/Lock/Key library

=Lock/Key library=

As mentioned in preliminary approaches in the section RNA-based Delay: Biosynthetic AND gate, our design was based on the previous work of Isaacs F. 2004, Berkeley iGEM 2006, Caltech iGEM 2007 and Peking iGEM 2007. However, we found that these projects were done using the "normal" RBS (B0034). The registry of biobricks has several RBS, besides, as a advice from our modeling team, the RBS present in our constructs should be one with low affinity or “weak” in order to avoid leakage problems. Therefore, we constructed an algorithm based on the previous works which allows us to generate pairs of locks and keys for any RBS (in or outside the registry).

Lock/Key library: Algorithm
Based on the various above mentioned studies on the locks and keys, we have developed an algorithm, which has led us to develop a simple web based tool to help in generating the DNA sequences of locks and keys for a chosen Ribosome Binding Site (RBS) DNA sequence. Click here for more information.

Lock/Key generator
You can also directly visit the web based Lock/Key generator page by clicking here.

Lock/Key library: New Biobricks Design
We want to test if the algorithm delivers functional locks and keys. In order to do that, we plan to characterize two locks and keys generated by our algorithm. The RBS’s chosen were the weak and medium (biobricks) from the registry. The DNA sequences of these new parts are... more

Results
Main results
 * Construction of an algorithm capable to generate locks (cis regulation) for different ribosome binding sites (RBS’s) and their correspond key (trans).
 * Online lock and key generator derived from the algorithm.
 * Construction of four DNA sequences in biobrick format and pSB1A3 plasmid backbone which encode lock for medium RBS (K175031), key for the lock of medium RBS (K175032), lock for weak RBS (K175029) or key for the lock of weak RBS (K175030).
 * Assembly of two composed biobricks (K175034 and K175035), designed to test the functionality of the lock/key pairs constructed.
 * Assembly of two GFP generators under the control of PLacI with weak RBS (K175033) or medium RBS (K175048)... more

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