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Developing a CRISPR-Mediated Knockout TCR Human T Cell Line for Use in Cloning Antigen-Specific T Cell Receptors


Abstract Adoptive transfer of T cells engineered to express synthetic antigen-specific T cell receptors (TCRs) has provocative therapeutic applications for treating cancer. However, expressing these synthetic TCRs in a CD4+ T cell line is a challenge. The CD4+ Jurkat T cell line expresses endogenous TCRs that compete for space, accessory proteins, and proliferative signaling, and there is the potential for mixed dimer formation between the α and β chains of the endogenous receptor and that of the synthetic cancer-specific TCRs. To prevent hybridization between the receptors and to ensure the binding affinity measured with flow cytometry analysis is between the tetramer and the TCR construct, a CRISPR-Cas9 gene editing pipeline was developed. The gui... (more)
Created Date 2020
Contributor Hirneise, Gabrielle Rachel (Author) / Anderson, Karen (Advisor) / Mason, Hugh (Committee member) / Lake, Douglas (Committee member) / Arizona State University (Publisher)
Subject Biology / Cellular biology / Cancer / CRISPR / Immunology / Immunotherapies / T cell / T cell receptor
Type Masters Thesis
Extent 65 pages
Language English
Copyright
Note Masters Thesis Biology 2020
Collaborating Institutions Graduate College / ASU Library
Additional Formats MODS / OAI Dublin Core / RIS


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Description Dissertation/Thesis