Skip to Main Content
 

Global Search Box

 
 
 
 

Files

File List


ucin1163452507.pdf (3.21 MB)

ETD Abstract Container

Abstract Header

THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS

LARSON, JON SCOTT
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507

Abstract Details

2006, PhD, University of Cincinnati, Medicine : Molecular Genetics, Biochemistry, and Microbiology.
Multicellular organisms are mosaic in nature because of genetic alterations that occur in somatic cells. There are many factors that can contribute to the formation of such alterations including aberrant DNA repair, environmental insults, epigenetic modification, errors in DNA replication and errors in chromosome duplication/segregation. To further the study of the distributions, frequencies and rates at which some alterations can occur, mouse reporter models were implemented. The Tg(âA-G11PLAP) transgenic mutation reporter mouse harbors an allele (G11 PLAP ) that is rendered incapable of producing its functional enzyme because of a reading frame shift caused by an insertion of 11 G:C basepairs. Spontaneous deletion of one G:C basepair from this mononucleotide repeat restores gene function, and cells with PLAP activity can be detected histochemically. G11 PLAP mice enable mutant cells to be visualized in situ and were used to study variation during early development, in the germline, under oxidative stress and in solid tumors. To study LOH in diverse cell types in the body another reporter model was implemented. Mice that carry two different fluorescent protein genes as alleles of a locus were generated to address this issue because LOH would change a cell’s phenotype from bichrome to monochrome. As a step in assessing the utility of this approach, we derived MEF and ES cell lines from mice that carried two different fluorescent protein genes as alleles at the chromosome 6 locus, ROSA26. FACS showed that the vast majority of cells in each line expressed the two marker proteins at similar levels, but populations exhibited extrinsic and intrinsic noise with respect to expression. In addition, cells with a monochrome phenotype were frequent (10-4). In ES cells, all monochrome events were accompanied by allele loss. Mitotic recombination appeared to be the major cause, although UPD also appeared to have contributed to LOH. These cells provided a novel assay for studying genetic/karyotypic stability of cultured ES cells. Results obtained from studies with these cells support the need for caution regarding the use of cultured stem cells in therapy.
James Stringer (Advisor)
141 p.
mutation; genomic instability; placental alkaline phospatase; microsatellite instability; loss of heterozygosity; embryonic stem cells; uniparental disomy; germline mutation; oxidative stress; maternal effect; harlequin mouse; mitotic recombination

Recommended Citations

Citations

  • LARSON, J. S. (2006). THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS [Doctoral dissertation, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507

    APA Style (7th edition)

  • LARSON, JON. THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS. 2006. University of Cincinnati, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507.

    MLA Style (8th edition)

  • LARSON, JON. "THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS." Doctoral dissertation, University of Cincinnati, 2006. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507

    Chicago Manual of Style (17th edition)

ucin1163452507
711
© 2006, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.