A novel disulphide containing phosphotriester monomer has been synthesised which possesses oligonucleotide coupling efficiencies which are superior to those obtained from existing disulphide monomers. This dipropyl disulphide linker, which should be stable outside the cell but vulnerable to disulphide cleavage by thiols contained in the cell, has been introduced between the lipophilic groups and oligonucleotides. The oligonucleotides should thus be released from the lipophilic groups once endocytosis has occurred. Similarly, the dipropyl disulphide linker has been introduced between a biotin group and an oligonucleotide. Quantitative disulphide cleavage can be achieved by the addition of dithiothreitol. Thus oligonucleotides bearing cleavable biotin moieties have been synthesised, employing a far more facile and high yielding process than the current method of choice. This approach has been extended to the synthesis of a novel amino functionalised disulphide linker for the production of the first cleavable aminolink oligonucleotides. Cleavage of the disulphide bond in the dipropyl disulphide linker produces a propyl thiol group at the 5'-end of an oligonucleotide. A diethyl disulphide linker has been synthesised which produces an ethyl thiol group on the oligonucleotide after disulphide cleavage. Under basic conditions, the oligonucleotide can eliminate mercaptoethanol to give a 5'-phosphate group. Thus a simple, high-yielding method for the synthesis of terminal thiol and terminal phosphate functionalised oligonucleotides has been developed, offering a viable alternative to existing routes to these functionalities.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:663788 |
| Date | January 1994 |
| Creators | Wilkinson, Adrian J. |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/11574 |
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