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The University of Southampton
Biological Sciences

Research project: Extending the boundaries of nucleic acid chemistry

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We are part of this BBSRC-funded sLoLa project, led by Prof Tom Brown (Chemistry), and are using click-chemistry to generate unusual DNA structures and examine their biological properties.

The click-chemistry methodology for ligating and assembling DNA fragments is being used to generate constructs that contain unusual DNA structures, such as G-quadruplexes and intramolecular triplexes, at unique positions and to assess their effects on transcription. Studies on these non-duplex structures in vivo are limited because of their transient nature, and the necessary presence of the complementary Watson-Crick strand, which enables them to revert to the familiar duplex conformation. The development of the click-ligation strategy will enable us to produce synthetic DNA plasmids which lack the complementary strands in these regions, thereby permanently fixing the otherwise transient structures. Synthetic plasmids can be prepared in large amounts, in which either the G- or C-rich strand is not paired with its complement, but is placed opposite a short loop, thereby forcing quadruplex formation. Similarly we will prepare synthetic plasmids which contain 5-methylcytosine (singly or multiply substituted, hemi- or fully-methylated). These will be used to assess the importance of individual methylation sites in epigenetic studies

This work is supported by BBSRC grant BB/J001694/1

Related research groups

Molecular and Cellular Biosciences
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