TY - JOUR T1 - Gene editing enables T-cell engineering to redirect antigen specificity for potent tumor rejection JF - Life Science Alliance JO - Life Sci. Alliance DO - 10.26508/lsa.201900367 VL - 2 IS - 2 SP - e201900367 AU - Julian J Albers AU - Tim Ammon AU - Dario Gosmann AU - Stefan Audehm AU - Silvia Thoene AU - Christof Winter AU - Ramona Secci AU - Anja Wolf AU - Anja Stelzl AU - Katja Steiger AU - Jürgen Ruland AU - Florian Bassermann AU - Christian Kupatt AU - Martina Anton AU - Angela M Krackhardt Y1 - 2019/04/01 UR - https://www.life-science-alliance.org/content/2/2/e201900367.abstract N2 - Adoptive transfer of TCR transgenic T cells holds great promise for treating various cancers. So far, mainly semi-randomly integrating vectors have been used to genetically modify T cells. These carry the risk of insertional mutagenesis, and the sole addition of an exogenous TCR potentially results in the mispairing of TCR chains with endogenous ones. Established approaches using nonviral vectors, such as transposons, already reduce the risk of insertional mutagenesis but have not accomplished site-specific integration. Here, we used CRISPR-Cas9 RNPs and adeno-associated virus 6 for gene targeting to deliver an engineered TCR gene specifically to the TCR alpha constant locus, thus placing it under endogenous transcriptional control. Our data demonstrate that this approach replaces the endogenous TCR, functionally redirects the edited T cells’ specificity in vitro, and facilitates potent tumor rejection in an in vivo xenograft model. ER -