Synthesis of light-activated antisense oligodeoxynucleotide.
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 (lambda approximately 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.
Tang
XinJing
X
Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, Pennsylvania 19104-6323, USA.
Dmochowski
Ivan J
IJ
D020319:Oligodeoxyribonucleotides, Antisense / Q000138:chemical synthesis* / Q000302:isolation & purification; D010782:Photolysis
D020319:Oligodeoxyribonucleotides, Antisense