Cross-reactive arrays based on three-way junctions.
We report herein a novel system for the parallel processing of molecular recognition events utilizing arrays of oligonucleotide-based fluorescent sensors to characterize hydrophobic molecules in solution. The binding domains of the sensors were based on three-way junctions that utilize double helical stems as framework regions to reliably fold regardless of variations in or around the binding domain. A reporting domain was introduced by the specific substitution of a single phosphodiester group with a phosphorothioate, followed by selective functionalization with a fluorophore. The sensors were organized into cross-reactive arrays to yield characteristic fingerprints for samples containing hydrophobic molecules. The fingerprints can be used to characterize steroids in solution, including complex biologically important fluids. Arrays have the potential for clinical applications such as the detection of gross errors in steroidogenesis.
Stojanović
Milan N
MN
Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, New York, New York 10032, USA. mns18@columbia.edu
Green
Eric G
EG
Semova
Stanka
S
Nikić
Dragan B
DB
Landry
Donald W
DW
D015374:Biosensing Techniques / Q000379:methods*; D003042:Cocaine / Q000032:analysis; D003429:Cross Reactions; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D010757:Phosphorous Acids / Q000138:chemical synthesis / Q000737:chemistry*; D013329:Structure-Activity Relationship
D005456:Fluorescent Dyes; D008024:Ligands; D009841:Oligonucleotides; D010757:Phosphorous Acids; D003042:Cocaine