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Title: DNA aptamers that selectively label eukaryotic cells depending on the expression of the cell surface protein, P2X7
Author: Stephens, Matthew Jon Craig
Awarding Body: University of Portsmouth
Current Institution: University of Portsmouth
Date of Award: 2013
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The ATP gated cation channel P2X7, is a cell surface protein whose function and activation is linked with several human diseases. The aim of this project was to raise DNA Aptamers targeting specifically the extracellular domain of the P2X7 protein with the hope to use these new ligands as a method for the study of the biological functions of the P2X7 protein and its potential as a therapeutic target. The strategy involved isolating Aptamers that would bind to human em~ryonic kidney cells transformed with a construct which forced the cells to express native mouse P2X7 on its cell surface (POSITIVE SELECTION), whilst not binding to untransformed non-P2X7 expressing cells (NEGATIVE SELECTION). Two candidate sequences from this selection emerged as, what we called, "winners" which were the most likely sequences to putatively bind mP2X7 (work ongoing). Interestingly using the procedure in reverse, that is to create Aptamers that specifically bind to non-transformed cells and not transformed cells, also produced viable putative Aptamers, which, once labelled, showed selective binding. These results are discussed in relation to future applications and the potential insights and implications for the study of cell surface protein expression patterns. 3
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available