Strain Promoted Click Chemistry of 8-Azidopurine and 5-Azidopyrimidine Nucleosides and Nucleotides with Cyclooctynes and Applications to Living Cell Imaging

Zayas, Jessica

doi:10.25148/etd.fidc000125

Cited by 1 publication

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References 101 publications

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“…We observed strong blue fluorescence in live cells using excitation and absorbance filters 360/40 and 470/40 nm, respectively (Figure 5), which was comparable to the fluorescence that Ito et al observed during the reaction of 8-azido-cAMP and a difluorinated cyclooctyne in HeLa cells. 46 However, contrary to this report, we found that the use of transfection agents (e.g., Lipofectamine; see SI for discussion), which often results in cytosol fluorescence, undue stress, and morphological changes in cells, was unnecessary.…”

Section: ■ Results and Discussioncontrasting

confidence: 74%

“…These prerequisites are, however, unsuitable for biological applications including the potential medicinal applications because of the harsh conditions used and cytotoxic effect of the copper catalyst. 45,46 Moreover, the coupling between 5-azidouridine analogues and terminal alkynes are also scarcely developed, 47 mainly because of the photochemical instability of the 5-azidouracil substrates. 48 To overcome these limitations, the 5-(azido)methyluracil nucleosides were used instead to study the click chemistry of azidomodified pyrimidine bases.…”

Section: ■ Introductionmentioning

confidence: 99%

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Strain Promoted Click Chemistry of 2- or 8-Azidopurine and 5-Azidopyrimidine Nucleosides and 8-Azidoadenosine Triphosphate with Cyclooctynes. Application to Living Cell Fluorescent Imaging

Zayas¹,

Annoual²,

Das³

et al. 2015

Bioconjugate Chem.

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Strain-promoted click chemistry of nucleosides and nucleotides with an azido group directly attached to the purine and pyrimidine rings with various cyclooctynes in aqueous solution at ambient temperature resulted in efficient formation (3 min - 3 h) of fluorescent, light-up, triazole products. The 2- and 8-azidoadenine nucleosides reacted with fused cyclopropyl cyclooctyne, dibenzylcyclooctyne or monofluorocyclooctyne to produce click products functionalized with hydroxyl, amino, N-hydroxysuccinimide, or biotin moieties. The 5-azidouridine and 5-azido-2′-deoxyuridine were similarly converted to the analogous triazole products in quantitative yields in less than 5 minutes. The 8-azido-ATP quantitatively afforded the triazole product with fused cyclopropyl cyclooctyne in aqueous acetonitrile (3 h). The novel triazole adducts at the 2 or 8 position of adenine or 5-position of uracil rings induce fluorescence properties which were used for direct imaging in MCF-7 cancer cells without the need for traditional fluorogenic reporters. FLIM of the triazole click adducts demonstrated their potential utility for dynamic measuring and tracking of signaling events inside single living cancer cells.

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Section: ■ Results and Discussioncontrasting

confidence: 74%