Prioritized selection of oligodeoxyribonucleotide probes for efficient hybridization to RNA transcripts.
Only a small fraction of short oligonucleotide probes bind efficiently to complementary segments in long RNA transcripts. Technologies such as array-based transcript profiling and antisense control of gene expression would benefit greatly from a method for predicting probes that bind well to a given target RNA. To develop an algorithm for prioritizing selection of probes, we have analyzed predicted thermodynamic parameters for the binding of several large sets of probes to complementary RNA transcripts. The binding of five of these sets of probes to their RNA targets has been reported by others. In addition, we have used a method for light-directed synthesis of oligonucleotide arrays that we developed to generate two new arrays of surface-bound probes and measured the binding of these probes to their RNA targets. We considered predicted free energies for intramolecular base pairing of the oligonucleotide and its RNA target as well as the predicted free energy of intermolecular hybridization of probe and target. We find that a reliable predictor of probes that will hybridize significantly with their targeted transcripts is the predicted free energy of hybridization minus the predicted free energy for intramolecular folding of the probe.
Luebke
Kevin J
KJ
Center for Biomedical Inventions, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390-9185, USA. kevin.luebke@utsouthwestern.edu
Balog
Robert P
RP
Garner
Harold R
HR
D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics*; D012313:RNA / Q000235:genetics*; D013816:Thermodynamics; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
D008164:Luminescent Proteins; D015345:Oligonucleotide Probes; D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins; D012313:RNA