The long-term goal of this laboratory is to use cellular and molecular methodologies to study the organization and expression of human chromosomes and their constituent genes and regulatory sequences in normal and disease states.
The specific aim of this component proposal is to use somatic cell genetics, molecular probes, microdissection and microcloning techniques for fine structure mapping, dissection and expression of human chromosome 21. These studies will provide direct link bridging cytogenetic and molecular resolution of chromosome 21 and can facilitate our search for genes and regulatory sequences underlying the etiology of Down syndrome. The proposed studies include (1) mapping of chromosome 21 using a series of well-defined terminal deletions, (2) microcloning of DNA sequences from defined areas of chromosome 21, and (3) combined use of microdissection and microinjection for studying expression of large blocks of DNA in cultured cells and animals. For terminal deletion mapping, we will establish large numbers of terminal deletions of chromosome 21 by using a human/CHO cell hybrid containing a 21q+ translocation chromosome derived from AML-M2 leukemia cells of t(8;21)(q22;q22.3). Terminal deletions will be produced by X-ray and selected by BrdU + visible light method using the auxotrophic marker GlyB which was translocated from chromosome 8 to 21 of the 21q+ translocation chromosome. Large numbers of DNA probes will be used to define the deletions in linear and submicroscopic resolution. These deletions will be used for refined regional mapping of all genes and DNA probes assigned to chromosome 21, and for resolving the order of many closely linked genes and DNA markers on this chromosome. For microdissecting chromosome 21, we will use the newly developed microdissection and microcloning techniques. Chromosome 21 can be subdivided into five regions and region-specific libraries will be constructed for providing DNA probes from defined areas of the chromosome for possible identification of DNA sequences associated with Down syndrome. Finally we will introduce large blocks of contiguous DNA sequences from dissected regions of chromosome 21 into cells by microinjection for studying their integration and expression. This work will be extended to transgenic mice. This approach appears particularly important for studying expression of complex disorders like Down syndrome.
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| Moat, Stuart; Carling, Rachel; Nix, Authur et al. (2010) Multicentre age-related reference intervals for cerebrospinal fluid serine concentrations: implications for the diagnosis and follow-up of serine biosynthesis disorders. Mol Genet Metab 101:149-52 |
| Nielsen, Darci M; Evans, Jeffrey J; Derber, William J et al. (2009) Mouse model of fragile X syndrome: behavioral and hormonal response to stressors. Behav Neurosci 123:677-86 |
| Knox, Aaron J; Graham, Christine; Bleskan, John et al. (2009) Mutations in the Chinese hamster ovary cell GART gene of de novo purine synthesis. Gene 429:23-30 |
| Hoger, Joachim; Patterson, David; Hoger, Harald et al. (2009) Mice transgenic for reduced folate carrier: an animal model of Down syndrome? Amino Acids 36:349-57 |
| Patterson, David; Graham, Christine; Cherian, Christina et al. (2008) A humanized mouse model for the reduced folate carrier. Mol Genet Metab 93:95-103 |
| Pennington, Bruce F (2006) From single to multiple deficit models of developmental disorders. Cognition 101:385-413 |
| Yao, Guimei; Chen, Xiao-Ning; Flores-Sarnat, Laura et al. (2006) Deletion of chromosome 21 disturbs human brain morphogenesis. Genet Med 8:1-7 |
| Wenger, Galen R; Schmidt, Cecilia; Davisson, Muriel T (2004) Operant conditioning in the Ts65Dn mouse: learning. Behav Genet 34:105-19 |
| Gardiner, Katheleen; Davisson, Muriel T; Crnic, Linda S (2004) Building protein interaction maps for Down's syndrome. Brief Funct Genomic Proteomic 3:142-56 |
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