The overall objective of this project is to develop and evaluate improved analytical cytologic techniques for prenatal detection of disease-linked cytogenetic abnormalities. Emphasis will be on a) improving the speed and sensitivity with which the loss or gain of chromosomal DNA can be detected and quantified during analysis of metaphase chromosomes, b) detection of common cytogenetic abnormalities in interphase cells and c) developing techniques for cytogenetic analysis using fetal cells isolated from maternal blood. These objectives will be pursued by: 1) Applying bivariate flow karyotyping to quantification of the extent of chromosome deletions and duplications as small as 1 Mb. Effort during this project period will be devoted to further increasing the sensitivity of the technique and applying it in a series of collaborative studies to characterize subtle structural aberrations of clinical or molecular biological importance. 2) Optimizing fluorescence in situ hybridization with chromosome specific probes for reliable detection and characterization of numerical aberrations involving chromosomes 21, 18, 13, x and y; segmental duplications (initially 21q22.3 for Down syndrome) and deletions (initially 15q11-13 for Prader-Willi syndrome) in metaphase spreads and in interphase nuclei. 3) Developing chromosome-specific probes to support the fluorescence hybridization studies. Whole-chromosome DNA and ssRNA probes depleted in repetitive sequences will be generated from chromosome-specific cosmid libraries. Composite probes for 21q22.3, 15q11-13 and elsewhere will be generated by starting with """"""""seed"""""""" probes already mapped to these regions and """"""""walking"""""""" outward in chromosome-specific cosmid libraries to expand the regions covered by the composite probes. 4) Continuing efforts to isolate fetal cells from maternal blood for interphase cytogenetic analysis. Specific projects include: a) determination of the gestational age within the first trimester at which the fetal cells are present at highest frequency in maternal blood,b)determination of the origin of the fetal cells (e.g. leukocytes or cytotrophoblasts) and c) development of procedures for enrichment of the fetal cells.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD017665-11
Application #
3314661
Study Section
Special Emphasis Panel (SSS)
Project Start
1991-07-01
Project End
1994-11-30
Budget Start
1991-12-01
Budget End
1992-11-30
Support Year
11
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Zhang, Xiaoxiao; Snijders, Antoine; Segraves, Richard et al. (2005) High-resolution mapping of genotype-phenotype relationships in cri du chat syndrome using array comparative genomic hybridization. Am J Hum Genet 76:312-26
Snijders, Antoine M; Segraves, Richard; Blackwood, Stephanie et al. (2004) BAC microarray-based comparative genomic hybridization. Methods Mol Biol 256:39-56
Albertson, Donna G (2003) Profiling breast cancer by array CGH. Breast Cancer Res Treat 78:289-98
Bruder, C E; Hirvela, C; Tapia-Paez, I et al. (2001) High resolution deletion analysis of constitutional DNA from neurofibromatosis type 2 (NF2) patients using microarray-CGH. Hum Mol Genet 10:271-82
Snijders, A M; Nowak, N; Segraves, R et al. (2001) Assembly of microarrays for genome-wide measurement of DNA copy number. Nat Genet 29:263-4
Albertson, D G; Ylstra, B; Segraves, R et al. (2000) Quantitative mapping of amplicon structure by array CGH identifies CYP24 as a candidate oncogene. Nat Genet 25:144-6
Pinkel, D; Segraves, R; Sudar, D et al. (1998) High resolution analysis of DNA copy number variation using comparative genomic hybridization to microarrays. Nat Genet 20:207-11
Yu, L C; Moore 2nd, D H; Magrane, G et al. (1997) Objective aneuploidy detection for fetal and neonatal screening using comparative genomic hybridization (CGH). Cytometry 28:191-7
Moore 2nd, D H; Pallavicini, M; Cher, M L et al. (1997) A t-statistic for objective interpretation of comparative genomic hybridization (CGH) profiles. Cytometry 28:183-90
Mascio, L N; Verbeek, P W; Sudar, D et al. (1995) Semiautomated DNA probe mapping using digital imaging microscopy: I. System development. Cytometry 19:51-9

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