Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals.

Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Research Abstract Details 

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Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Abstract Text:

C Anthony Blau,Kenneth R Peterson,

Transgenic mice produced with human yeast artificial chromosomes (YACs) generally display transgene expression patterns that reflect those of the normal human host. Because mice are expensive and time-consuming to generate and maintain, extensive mutation-phenotype correlation studies cannot be readily carried out. Cell lines are better suited for analysis of a plethora of mutations. However, these types of gene regulatory studies have been complicated by the lack of suitable cell lines, most of which do not exactly replicate the gene expression patterns observed in vivo. We reasoned that cells established from tissues of YAC transgenic mice might express the transgenes in the correct tissue and developmental stage-specific pattern from which they were derived because YAC transgenic mice display correct regulation of gene expression during ontogeny. We used our human beta-globin locus YAC (beta-YAC) transgenic mice to demonstrate this approach. All existing erythroid cell lines coexpress beta-like globins from different developmental stages or express them inappropriately based on the developmental stage from which they were obtained. Cell populations were established from the adult bone marrow (BM) of beta-YAC transgenic mice, which express exclusively adult beta-globin, using dimerizer technology. A derivative of the thrombopoietin receptor (mpl) was used to bring the proliferative status of primary BM marrow cells under the control of a small molecule drug called a chemical inducer of dimerization (CID). Cells generated in this manner can be expanded to extremely large numbers, remain strictly CID-dependent, and retain megakaryocytic, erythroid, and granulocytic potential. Marrow cells transduced with a retrovirus vector encoding the mpl derivative proliferated extensively in the presence of the CID, AP20187. RNAse protection assays demonstrated that the transcripts for human beta-globin and mouse alpha-globin were present, while gamma-globin transcripts were absent, thus, these cells had the predicted expression phenotype. Exposure to 5-azacytidine or introduction of a hereditary persistence of fetal hemoglobin mutation activated gamma-globin, which was expressed in addition to beta-globin, again consistent with the predicted expression profile of these cells. This approach extends the usefulness of YAC transgenic mice for the generation of cell lines amenable to more detailed studies regarding gene regulation.

Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Publishing Authors By Initials

CA Blau,KR Peterson,

For similar enzymes and coenzymes: enzymes: isomerases: cis-trans-isomerases: peptidylprolyl isomerase: immunophilins: tacrolimus binding proteins research abstracts see: enzymes and coenzymes: enzymes: isomerases: cis-trans-isomerases: peptidylprolyl isomerase: immunophilins: tacrolimus binding proteins research

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Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Methods in molecular biology (Clifton, N.J.)

VOLUME: 349

Page Numbers: 163-73

Journal Abbreviation: Methods Mol. Biol.

ISSN: 1064-3745

DAY: 3

MONTH: 12

YEAR: 2006

Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9214969

Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Keywords Mesh Terms:

KEYWORDS: Tacrolimus Binding Proteins

MESH TERMS: chemistry

Chemical & Substance for Abstract: Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals. Information

Substance Name: Tacrolimus Binding Proteins

Registry Number: EC 5.2.1.-

Grant and Affiliation Information for Establishment of cell lines that exhibit correct ontogenic stage-specific gene expression profiles from tissues of yeast artificial chromosome transgenic mice using chemically induced growth signals.

AFFILIATION: Division of Hematology, University of Washington, Seattle, USA.

Country: United States

AGENCY: United States NHLBI

GRANT: HL67336

ACRONYM: HL

MEDLINETA: Methods Mol Biol

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