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Title: Constructing RNA based genetic circuits for the detection of endogenous microRNAs
Author: Rajakumar, Timothy
ISNI:       0000 0004 7234 1584
Awarding Body: University of Oxford
Current Institution: University of Oxford
Date of Award: 2016
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Synthetic biology is a rapidly evolving field enabling the rational design of new biological functionality. RNA is particularly amenable to this rational design processes due to relatively straightforward structural modelling principles and its ability to perform diverse roles, from that of simple messenger RNAs to complex catalytic ribozymes. Consequently, the synthetic design and assembly of RNA building blocks into functional circuits and networks can far exceed the complexity of what could be achieved using protein components. MicroRNAs (miRNAs) are fascinating short non-coding RNA molecules (~22nt long) which have the capacity to bind and tune the expression of thousands of mRNAs, functioning as molecular rheostats during development, physiology and disease. The ability to detect miRNAs, via synthetic biosensor circuits would therefore offer a unique insight into cellular behavior in addition to the ability to link miRNA expression signatures to an output expression programme of choice. In this thesis, I have developed novel synthetic circuit architectures for the detection of miRNA activity. Firstly, protein and RNA modules were used in concert to develop a biosensor circuit consisting of two serially linked NOT gates that could activate output expression in response to a specific input miRNA. Secondly, methods were devised for the construction of a single molecule biosensor device, built entirely using RNA. At the heart of this device were novel miRNA responsive allosteric ribozymes which have been created through two independent in vitro selection methodologies devised for this study. Such engineered modules now constitute part of the synthetic biology toolbox, increasing the versatility with which miRNAs can serve as inputs to artificial genetic circuits.
Supervisor: Fulga, Tudor Sponsor: MRC
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available