Author: Banu, Nasirah; Chia, Adeline; Ho, Zi Zong; Garcia, Alfonso Tan; Paravasivam, Komathi; Grotenbreg, Gijsbert M.; Bertoletti, Antonio; Gehring, Adam J.
Title: Building and Optimizing a Virus-specific T Cell Receptor Library for Targeted Immunotherapy in Viral Infections Document date: 2014_2_25
ID: 44w6omdp_6
Snippet: Optimizing virus-specific TCR expression in primary human T cells. The wild type alpha and beta TCR pair was originally tested for expression by co-transducing two separate vectors carrying the TCR alpha and beta genes ( Fig. 1A; a 1 b) . To begin optimizing TCR expression, we made stepwise advancements in the TCR constructs based on published literature using 3 different virus-specific TCRs. The TCRs were linked together with a 2A autocleavage s.....
Document: Optimizing virus-specific TCR expression in primary human T cells. The wild type alpha and beta TCR pair was originally tested for expression by co-transducing two separate vectors carrying the TCR alpha and beta genes ( Fig. 1A; a 1 b) . To begin optimizing TCR expression, we made stepwise advancements in the TCR constructs based on published literature using 3 different virus-specific TCRs. The TCRs were linked together with a 2A autocleavage site and flexible spacer 13, 14 to promote equimolar production of the alpha and beta chains and codon optimized to increase translational efficiency 17 . These constructs were then modified with a nonnative cysteine disulfide bond in the constant region to increase pairing and stability of the exogenous TCR 15, 16 .
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