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Exceptional Chromosome Regions Workshop I

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Nucleic Acid Strands Specific for Binding via Non-Denaturing Triple Helix Formation (TISH) to Centromeric DNAs of Each of the Human Chromosomes and to the DNA Common to the Tips of the Achrocentrics

Jacques R. Fresco
Dept. of Molecular Biology, Princeton University, Princeton, N. J. 08544

Marion Johnson and I have developed a methodology for binding single nucleic acid strands specifically to purine-rich.pyrimidine-rich sequences of protein-depleted chromosomes under conditions that are non-denaturing to double helical DNA. The binding occurs by triple helix formation and is remarkably sequence-specific under conditions of appropriate stringency. The probe strands are 15-25 residues in length, can distinguish a single base pair difference in the target sequence, the binding is quantitative, and the method should be relatively easy to adapt for the isolation of a long DNA chain in which a target is present. So far we have used this approach to develop fluorescent cytogenetic probes in human and drosophila chromosomes (cf. M. D. Johnson and J. R. Fresco, CHROMOSOMA (1999) 108:181-189.Currently, we are using it as the basis of a novel approach to gene therapy and also to develop cytogenetic probes for genes amplified in various types of Cancer.

My laboratory will be pleased to collaborate, help, or otherwise interact with anyone interested in exploiting this technology.


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