TARGETING OF SELECTABLE MARKERS TO ANY GENETIC LOCUS

IT IS THE LONG-TERM GOAL OF THIS RESEARCH TO DEVELOP A METHOD OF ORDERLY SEGMENTING THE HUMAN GENOME INTO 1-50 MB FRAGMENTS. THIS OBJECTIVE CAN BE ACHIEVED BY STREAMLINING THE SCREENING PROCESS OF HYBRID CLONES GENERATED BY CHROMOSOME FRAGMENT GENE TRANSFER. THE MOST EXPEDITIOUS SCREENING PROCESS IS ONE THAT INCLUDES A DOMINANT SELECTABLE MARKER NEAR THE LOCUS OF INTEREST. HOMOLOGOUS RECOMBINATION WILL BE USED TO DIRECT A DOMINANT SELECTABLE MARKER TO A SPECIFIC CHROMOSOMAL LOCATION ALLOWING THE TARGETED LOCUS TO BECOME PHENOTYPICALLY SELECTABLE. CELLS CONTAINING A CORRECTLY TARGETED DOMINANT MARKER ARE AN IDEALDONOR SOURCE FOR GOSS-HARRIS HYBRIDS. THE TRANSGENOMES PRODUCED BY THIS APPROACH WILL BE VALUABLE REAGENTS FOR MOLECULAR AND MEDICAL GENETIC RESEARCH. THEY WILL ALLOW FOR A RAPID MEANS OF SATURATING A LOCUS WITH GENETIC MARKERS. IN ADDITION, THE RESULTING ENRICHMENT OF ATARGETED GENETIC DISEASE LOCUS WILL FACILITATE THE IDENTIFICATION OF THE RESPONSIBLE DEFECT. AT THE GENOME LEVEL, A SET OF TARGETING VECTORS, EACH DIRECTED TO CHROMOSOMAL LOCATIONS SEPARATED BY 10-20 MB, COULD GENERATE AN ORDERED SET OF TRANSGENOMES. THESE REAGENTS WOULD ENHANCE CURRENT AND FUTURE HUMAN GENOME MAPPING EFFORTS.