Open AccessSplice-Switching Antisense Oligonucleotides as a Targeted Intrinsic Engineering Tool for Generating Armored Redirected T Cells
Author(s) -
Erica Ceccarello,
Tommaso Tabaglio,
Sarene Koh,
Vincent Yi Sheng Oei,
Winnie Koon Lay Teo,
Owen Julianto Jonathan,
Andrea Pavesi,
YiPing Phoebe Chen,
Antonio Bertoletti,
Keng Boon Wee,
Ernesto Guccione
Publication year - 2021
Publication title -
nucleic acid therapeutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.255
H-Index - 67
eISSN - 2159-3345
pISSN - 2159-3337
DOI - 10.1089/nat.2020.0905
Subject(s) - electroporation , transfection , oligonucleotide , t cell , microbiology and biotechnology , t cell receptor , exon , gene knockdown , exon skipping , biology , morpholino , computational biology , immune system , alternative splicing , cell culture , immunology , genetics , gene
Modification of specificity of T cells for the use in adoptive transfer (CAR- or TCR-redirected T cells) has revolutionized the therapy of liquid tumors and some infectious diseases. However, several obstacles are still hampering the efficacy of such potent therapy, hence concurrent modification of the function is also required to obtain successful results. Here we show the use of splice-switching antisense oligonucleotides (SSOs) as a tool to transiently modify T cell function. We demonstrate the possibility to transfect SSOs and an exogenous TCR into primary human T cells in the same electroporation reaction, without affecting viability and function of the transfected T lymphocytes. Moreover, we show that SSOs targeting T cell-specific mRNAs induce the skipping of the targeted exons, and the reduction of the protein and consequent modification of T cell function. This technical work paves the way to the use of SSOs in immune cells, not only for the knockdown of the functional isoform of the targeted proteins, but also for the protein manipulation by elimination of specific domains encoded by targeted exons.