Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above
The instability of DNA triplexes particularly at neutral pH and above severely limits their applications. Here, we demonstrate that the introduction of a thiazole orange (TO) intercalator onto a thymine nucleobase in triplex forming oligonucleotides (TFOs) resolves this problem. The stabilising effe...
| Główni autorzy: | , , |
|---|---|
| Format: | Journal article |
| Wydane: |
Royal Society of Chemistry
2018
|
| _version_ | 1826299247020998656 |
|---|---|
| author | Walsh, S El-Sagheer, A Brown, T |
| author_facet | Walsh, S El-Sagheer, A Brown, T |
| author_sort | Walsh, S |
| collection | OXFORD |
| description | The instability of DNA triplexes particularly at neutral pH and above severely limits their applications. Here, we demonstrate that the introduction of a thiazole orange (TO) intercalator onto a thymine nucleobase in triplex forming oligonucleotides (TFOs) resolves this problem. The stabilising effects are additive; multiple TO units produce nanomolar duplex binding and triplex stability can surpass that of the underlying duplex. In one example, a TFO containing three TO units increased the triplex melting temperature at pH 7 by a remarkable 50 °C relative to the unmodified triplex. Notably, TO intercalation promotes TFO binding to target sequences other than pure polypurine tracts by the use of 5-(1-propynyl)cytosine (pC) against C:G inversions. By overcoming the instability of triplexes across a broad range of pH and sequence contexts, these very simple ‘TOTFO’ probes could expand triplex applications into many areas including diagnostics and cell imaging. |
| first_indexed | 2024-03-07T04:59:00Z |
| format | Journal article |
| id | oxford-uuid:d79cbd11-6811-4778-9e7c-ff2743d78d39 |
| institution | University of Oxford |
| last_indexed | 2024-03-07T04:59:00Z |
| publishDate | 2018 |
| publisher | Royal Society of Chemistry |
| record_format | dspace |
| spelling | oxford-uuid:d79cbd11-6811-4778-9e7c-ff2743d78d392022-03-27T08:42:25ZFluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and aboveJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:d79cbd11-6811-4778-9e7c-ff2743d78d39Symplectic Elements at OxfordRoyal Society of Chemistry2018Walsh, SEl-Sagheer, ABrown, TThe instability of DNA triplexes particularly at neutral pH and above severely limits their applications. Here, we demonstrate that the introduction of a thiazole orange (TO) intercalator onto a thymine nucleobase in triplex forming oligonucleotides (TFOs) resolves this problem. The stabilising effects are additive; multiple TO units produce nanomolar duplex binding and triplex stability can surpass that of the underlying duplex. In one example, a TFO containing three TO units increased the triplex melting temperature at pH 7 by a remarkable 50 °C relative to the unmodified triplex. Notably, TO intercalation promotes TFO binding to target sequences other than pure polypurine tracts by the use of 5-(1-propynyl)cytosine (pC) against C:G inversions. By overcoming the instability of triplexes across a broad range of pH and sequence contexts, these very simple ‘TOTFO’ probes could expand triplex applications into many areas including diagnostics and cell imaging. |
| spellingShingle | Walsh, S El-Sagheer, A Brown, T Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above |
| title | Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above |
| title_full | Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above |
| title_fullStr | Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above |
| title_full_unstemmed | Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above |
| title_short | Fluorogenic thiazole orange TOTFO probes stabilise parallel DNA triplexes at pH 7 and above |
| title_sort | fluorogenic thiazole orange totfo probes stabilise parallel dna triplexes at ph 7 and above |
| work_keys_str_mv | AT walshs fluorogenicthiazoleorangetotfoprobesstabiliseparalleldnatriplexesatph7andabove AT elsagheera fluorogenicthiazoleorangetotfoprobesstabiliseparalleldnatriplexesatph7andabove AT brownt fluorogenicthiazoleorangetotfoprobesstabiliseparalleldnatriplexesatph7andabove |