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Developing a CRISPR-Mediated Knockou...

Hirneise, Gabrielle.

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

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Developing a CRISPR-Mediated Knockout TCR Human T Cell Line for Use in Cloning Antigen-specific T Cell Receptors./
作者:	Hirneise, Gabrielle.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2020,
面頁冊數:	66 p.
附註:	Source: Masters Abstracts International, Volume: 81-11.
Contained By:	Masters Abstracts International81-11.
標題:	Biology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27994689
ISBN:	9798645462864

Developing a CRISPR-Mediated Knockout TCR Human T Cell Line for Use in Cloning Antigen-specific T Cell Receptors.
Hirneise, Gabrielle.

Developing a CRISPR-Mediated Knockout TCR Human T Cell Line for Use in Cloning Antigen-specific T Cell Receptors. - Ann Arbor : ProQuest Dissertations & Theses, 2020 - 66 p.

Source: Masters Abstracts International, Volume: 81-11.

Thesis (M.S.)--Arizona State University, 2020.

This item must not be sold to any third party vendors.

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 guide RNAs (gRNAs) within the complex were designed to target the constant region of the α and β chains, as they are conserved between TCR clonotypes. To minimize further interference and confer cytotoxic capabilities, gRNAs were designed to target the CD4 coreceptor, and the CD8 coreceptor was delivered in a mammalian expression vector. Further, Golden Gate cloning methods were validated in integrating the gRNAs into a CRISPR-compatible mammalian expression vector. These constructs were transfected via electroporation into CD4+ Jurkat T cells to create a CD8+ knockout TCR Jurkat cell line for broadly applicable uses in T cell immunotherapies.

ISBN: 9798645462864Subjects--Topical Terms:

522710
Biology.
Subjects--Index Terms:

Cancer

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