Abstract
The activity of a 20-mer antisense oligodeoxynucleotide (asODN) is transiently blocked by attaching a partially complementary sense strand (sODN) via a heterobifunctional photocleavable linker (PL). The asODN-PL-sODN conjugate forms a DNA hairpin-like structure that is considerably more stable than the corresponding asODN/sODN duplex. In conjugate form, the asODN is prevented from hybridizing to exogenous RNA or DNA molecules. Activity is restored after modest exposure to UV light (λ ≈ 365 nm). Here, we provide a detailed procedure for synthesizing photoactive asODNs in good yields. Synthesis, purification and analysis of the light-activated asODN can be completed within 1–2 weeks.
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Acknowledgements
Support for this work came from grants from the Penn Genomics Institute (PGI) and Penn Institute for Medicine and Engineering (IME), and startup funds provided to I.J.D. by the Department of Chemistry at the University of Pennsylvania.
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Tang, X., Dmochowski, I. Synthesis of light-activated antisense oligodeoxynucleotide. Nat Protoc 1, 3041–3048 (2006). https://doi.org/10.1038/nprot.2006.462
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DOI: https://doi.org/10.1038/nprot.2006.462
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