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In vitro evolution of RNA ligands: understanding the process and exploiting the products

Reference	G10146
Principal Investigator / Supervisor	Professor William James
Co-Investigators / Co-Supervisors
Institution	University of Oxford
Department	Sir William Dunn Sch of Pathology
Funding type	Research
Value (£)	188,360
Status	Completed
Type	Research Grant
Start date	01/10/1998
End date	01/10/2001
Duration	36 months

Abstract

The process of in vitro evolution of nucleic acid ligands (aptamers) makes possible the development of analytical, diagnostic and therapeutic reagents in fields of study that have previously been refractory. For example, aptamers can be generated with specificity for non-immunogenic molecules, for highly conserved proteins and for rare structural variants, making use of the power of the pseudo-genetic process. However, the paucity of downstream technologies for exploitation and a surprising lack of understanding of the selection process itself have seriously limited the potential of the method. Here, we propose to build on our recent advances in both areas to make aptamers a generally useful tool. Using BIAcore methods, we have undertaken real-time kinetic analyses of RNAs at various cycles during the SELEX process, and these results have caused us to re-evaluate the standard methodology. We will develop the insights we have gained into the kinetic properties of the evolving aptamer population to gain a fuller understanding of the potential of dissociation-rate based selection in the generation of aptamers with high affinity and a diversity of specificities. We will devise methods for improved RNA refolding methods, which will enhance the power of each selection cycle. We will study a range of methods for producing useful homodimeric and heterodimeric aptamers and for tagging them with biotin and other labels. These advances will greatly increase the general utility of the methodology and be of benefit to UK science and business.

Summary

unavailable

Committee	Closed Committee - Genes & Developmental Biology (GDB)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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