The goal of our research is to dissect, understand, and control the biology of single cells in complex tissues, such as brain, or in malignant tumors. Furthering this body of work requires that we address a problem in single cell molecular analysis: the lack of a method to routinely, reliably and inexpensively determine global gene transcriptional activity.To solve this problem we aim to directly identify individual cDNA gene transcript molecules via atomic force microscopy (AFM). Each molecule is 'coded' by in situ digestion with sequence-specific restriction enzymes which produce detectable breaks in individual polynucleotide backbones, whose locations correspond to particular enzyme recognition sequences.  The advantage of this approach stems from: (1) the chemistry involves many fewer biochemical steps than do the alternatives, and (2) AFM's single molecule sensitivity, which allows enzymatic amplification to be eliminated. These improvements will significantly reduce time, cost and complexity of small sample transcriptional profiling.

Single molecule gene expression profiling using atomic force microscopy

241st National Meeting (2011)

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