In vitro models to study the properties of the tcralpha locus control region
Item
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Title
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In vitro models to study the properties of the tcralpha locus control region
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Identifier
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d_2009_2013:a08c3b27d3a5:12044
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identifier
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12747
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Creator
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Lahiji, Armin,
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Contributor
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Benjamin D. Ortiz
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Date
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2013
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Language
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English
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Publisher
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City University of New York.
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Subject
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Immunology | Biology | Genetics | Barroers | Development | Embryonic Stem Cells | Epigenetics | Insulators | Locus Control Regions
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Abstract
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Locus Control Regions (LCR) are cis-acting gene regulatory elements with the unique, integration site-independent ability to transfer the characteristics of their locus-of-origin's gene expression pattern to a linked transgene. LCR activities have been discovered in numerous T cell-lineage gene loci. These elements can be adapted to the design of gene therapy vectors that direct robust therapeutic gene expression to the T cell progeny of engineered stem cells. Currently, transgenic mice provide the only experimental approach that wholly supports all the critical aspects of LCR activity. Herein, we report two cell culture models to study the properties of the T cell receptor (TCR)-&agr; LCR. The first is an in vitro embryonic stem cell differentiation model that has been optimized to manifest all key features of mouse TCR&agr; LCR function. High level, copy number-related TCR&agr; LCR-linked reporter gene expression levels are cell type-restricted in this system, and are upregulated during the expected stage transition of T cell development. The ability of this LCR to overcome position effects may be due to barrier insulator-like activity within. To further explore this possibility, we established a second model that seems to support this notion. The characterization and identification of the sequences involved in this possible barrier insulator will provide an additional vertebrate model for the study of insulators. This study additionally validates a novel, tractable and more rapid approach for the study of LCR activity in T cells, and its translation to therapeutic genetic engineering.
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Type
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dissertation
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Source
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2009_2013.csv
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degree
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Ph.D.
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Program
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Biology
