東華大學圖書館 |

Language: English

Help

回圖書館首頁

手機版館藏查詢

Back

Switch To: Labeled | MARC Mode | ISBD

Developing a CRISPR-Mediated Knockou...

Hirneise, Gabrielle.

Linked to FindBook

Google Book

Amazon

博客來

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

Record Type:	Electronic resources : Monograph/item
Title/Author:	Developing a CRISPR-Mediated Knockout TCR Human T Cell Line for Use in Cloning Antigen-specific T Cell Receptors./
Author:	Hirneise, Gabrielle.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2020,
Description:	66 p.
Notes:	Source: Masters Abstracts International, Volume: 81-11.
Contained By:	Masters Abstracts International81-11.
Subject:	Biology. -
Online resource:	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.
LDR:02550nmm a2200373 4500 001 2270294
005 20200921070815.5
008 220629s2020 ||||||||||||||||| ||eng d
020 $a 9798645462864
035 $a (MiAaPQ)AAI27994689
035 $a AAI27994689
040 $a MiAaPQ $c MiAaPQ
100 1 $a Hirneise, Gabrielle. $3 3547667
245 1 0 $a Developing a CRISPR-Mediated Knockout TCR Human T Cell Line for Use in Cloning Antigen-specific T Cell Receptors.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2020
300 $a 66 p.
500 $a Source: Masters Abstracts International, Volume: 81-11.
500 $a Advisor: Anderson, Karen.
502 $a Thesis (M.S.)--Arizona State University, 2020.
506 $a This item must not be sold to any third party vendors.
520 $a 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.
590 $a School code: 0010.
650 4 $a Biology. $3 522710
650 4 $a Cellular biology. $3 3172791
653 $a Cancer
653 $a CRISPR
653 $a Immunology
653 $a Immunotherapies
653 $a T cell
653 $a T cell receptor
690 $a 0306
690 $a 0379
710 2 $a Arizona State University. $b Biology. $3 1673500
773 0 $t Masters Abstracts International $g 81-11.
790 $a 0010
791 $a M.S.
792 $a 2020
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27994689