THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS
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| Language: | English |
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University of Cincinnati / OhioLINK
2006
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| Online Access: | http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507 |
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ndltd-OhioLink-oai-etd.ohiolink.edu-ucin11634525072021-08-03T06:11:26Z THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS LARSON, JON SCOTT 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 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. 2006 English text University of Cincinnati / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507 http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws. |
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NDLTD |
| language |
English |
| sources |
NDLTD |
| topic |
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 |
| spellingShingle |
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 LARSON, JON SCOTT THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS |
| author |
LARSON, JON SCOTT |
| author_facet |
LARSON, JON SCOTT |
| author_sort |
LARSON, JON SCOTT |
| title |
THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS |
| title_short |
THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS |
| title_full |
THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS |
| title_fullStr |
THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS |
| title_full_unstemmed |
THE VISUALIZATION, QUANTIFICATION AND MODELING OF GENOMIC INSTABILITY IN THE MOUSE AND IN CULTURED CELLS |
| title_sort |
visualization, quantification and modeling of genomic instability in the mouse and in cultured cells |
| publisher |
University of Cincinnati / OhioLINK |
| publishDate |
2006 |
| url |
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1163452507 |
| work_keys_str_mv |
AT larsonjonscott thevisualizationquantificationandmodelingofgenomicinstabilityinthemouseandinculturedcells AT larsonjonscott visualizationquantificationandmodelingofgenomicinstabilityinthemouseandinculturedcells |
| _version_ |
1719432468650524672 |