The principle of the proposed project is the application of fluorescent microsphere based suspension array technology (SAT) to the cytometry based simultaneous detection of multi analytes. The proposal consists of four overlapping subprojects: (a) Technological development of biological microbead procedures using cells or microorganisms as substitutes for polymeric microspheres to lower the cost of SAT compared to the cost of methods using microbeads manufactured by polymer chemistry. (b) Development of a test to detect gene translocations and rearragements frequently responsible for malignant transformations. A method based on measuring bead-based hybridization of specific 5' and 3' nucleotide sequences in the vicinity of chromosomal translocation breakpoints would allow large scale screening of leukemias and various types of cancers by the measurement of 5'/3' sequence ratio. (c) Development of a DNA binding protein profiling assay providing easy, reproducible and high throughput technology to determine the expression pattern of transcription factors (TFs) binding to single stranded hexamere oligonucleotide sequences anchored to the solid phase surface of microspheres. This subproject is at the R&D level and needs to be thoroughly tested. (d) Development of artificial neural network software applications to analyze and classify the "fingerprint patterns" produced by the SAT measurements in subprojects (b) and (c). One of the most important goals of the work in Phase I will be to select the approach worth focusing on in Phase II.
Thesaurus Terms: DNA binding protein, bacteria, functional genomics, gene rearrangement, genome, microarray technology, polymer, technology /technique development, tissue /cell culture artificial intelligence, chromosome translocation, genetic recombination, high throughput technology, oligonucleotide, transcription factor flow cytometry
