Facile synthesis of advanced photodynamic molecular beacon architectures.
Nucleic acid photodynamic molecular beacons (PMBs) are a class of activatable photosensitizers that increase singlet oxygen generation upon binding a specific target sequence. Normally, PMBs are functionalized with multiple solution-phase labeling and purification steps. Here, we make use of a flexible solid-phase approach for completely automated synthesis of PMBs. This enabled the creation of a new type of molecular beacon that uses a linear superquencher architecture. The 3' terminus was labeled with a photosensitizer by generating pyropheophorbide-labeled solid-phase support. The 5' terminus was labeled with up to three consecutive additions of a dark quencher phosphoramidite. These photosensitizing and quenching moieties were stable in the harsh DNA synthesis environment and their hydrophobicity facilitated PMB purification by HPLC. Linear superquenchers exhibited highly efficient quenching. This fully automated synthesis method simplifies not only the synthesis and purification of PMBs, but also the creation of new activatable photosensitizer designs.
Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, Ontario.
Jonathan F
JF
Lovell
Juan
J
Chen
Elizabeth
E
Huynh
Mark T
MT
Jarvi
Brian C
BC
Wilson
Gang
G
Zheng
D001331:Automation; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000138:chemical synthesis / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D015335:Molecular Probes / Q000138:chemical synthesis* / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry; D017319:Photosensitizing Agents / Q000138:chemical synthesis* / Q000737:chemistry*; D026082:Singlet Oxygen / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D012997:Solvents / Q000737:chemistry
D015335:Molecular Probes; D009943:Organophosphorus Compounds; D017319:Photosensitizing Agents; D012996:Solutions; D012997:Solvents; C434331:phosphoramidite; D026082:Singlet Oxygen; D004247:DNA