Single molecule analysis of DNA/protein interactions: The nanopore shift assay

ANYL 247

Meni Wanunu, wanunu@bu.edu1, Devora Cohen-Karni, devorack@bu.edu1, Yong Yu1, Zhiping Weng1, and Amit Meller, ameller@bu.edu2. (1) Biomedical Engineering, Boston University, 44 Cummington St, Boston, MA 02215, (2) Department of Biomedical Engineering, Boston University, 44 Cummington St, Boston, MA 02215
Transcription factors (TFs) are an important class of gene regulatory proteins. They recognize short doubled stranded DNA stretches (up to 20 bp) with high specificity and affinity. We have recently developed a novel approach for analyzing DNA/TF interactions at the single-molecule level. Our method circumvents some of the shortcomings of common bulk approaches (gel shift assay and chromatin immunoprecipitation with microarray detection). It utilizes solid-state nanopores comparable in size to double-stranded DNA cross-section, to analyze TF binding with high throughput. TF-bound and unbound DNA molecules display distinct differences in the measured ion current patterns, enabling ultra fast detection and screening of TF binding. We present results demonstrating the feasibility of our approach using the TF SP1, which contain zinc finger motif, and map its interactions, with genomic DNA fragments. Future studies will allow us to study interactions between multiple TFs and neighboring binding sites in a single experiment.
 

General Posters
7:00 PM-9:00 PM, Sunday, August 19, 2007 BCEC -- Exhibit Hall - B2, Poster

Sci-Mix
8:00 PM-10:00 PM, Monday, August 20, 2007 BCEC -- Exhibit Hall - B2, Sci-Mix

Division of Analytical Chemistry

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007