Quantitative modeling and data analysis of SELEX experiments.
SELEX (systematic evolution of ligands by exponential enrichment) is an experimental procedure that allows the extraction, from an initially random pool of DNA, of those oligomers with high affinity for a given DNA-binding protein. We address what is a suitable experimental and computational procedure to infer parameters of transcription factor-DNA interaction from SELEX experiments. To answer this, we use a biophysical model of transcription factor-DNA interactions to quantitatively model SELEX. We show that a standard procedure is unsuitable for obtaining accurate interaction parameters. However, we theoretically show that a modified experiment in which chemical potential is fixed through different rounds of the experiment allows robust generation of an appropriate dataset. Based on our quantitative model, we propose a novel bioinformatic method of data analysis for such a modified experiment and apply it to extract the interaction parameters for a mammalian transcription factor CTF/NFI. From a practical point of view, our method results in a significantly improved false positive/false negative trade-off, as compared to both the standard information theory based method and a widely used empirically formulated procedure.
Djordjevic
Marko
M
Department of Physics, Columbia University, New York, NY 10027, USA. mdjordjevic@math.ohio-state.edu
Sengupta
Anirvan M
AM
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003198:Computer Simulation; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005786:Gene Expression Regulation; D008024:Ligands; D008954:Models, Biological; D008969:Molecular Sequence Data; D051839:NFI Transcription Factors / Q000737:chemistry / Q000378:metabolism; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D052156:SELEX Aptamer Technique
D008024:Ligands; D051839:NFI Transcription Factors; D004247:DNA