Adeno-Associated Viral Vectors for Homology-Directed Generation of CAR-T Cells.
Immunotherapy with T cells expressing chimeric antigen receptors (CAR) is an emerging and promising treatment against refractory cancers. However, the currently adopted methods of modification of T cells pose a risk of insertional oncogenesis because lentiviral and retroviral vectors integrate the CAR transgene in a semi-random fashion. In addition, this therapy is only available using autologous cells, which create problems in production and limit the access for patients who have their T cells depleted. One modification method that shows the ability to overcome both drawbacks is the knock-in of the CAR simultaneously knocking-out genes that prevent allogeneic therapy, such as the endogenous T cell receptor. In this mini-review, the authors present recent efforts to develop safer universal CAR-T cells. More specifically, the combined application of target-directed nucleases, which create a double-strand break at a specific genome locus, and the delivery of CAR DNA via adeno-associated viral vectors for subsequent integration via homologous recombination and silencing of the targeted gene is focused on.
['Moço|Pablo D|PD|https://orcid.org/0000-0001-9405-4242', 'Aharony|Noga|N|', 'Kamen|Amine|A|https://orcid.org/0000-0001-9110-8815']
D000229:Dependovirus D004720:Endonucleases D000072669:Gene Editing D005822:Genetic Vectors D006801:Humans D016219:Immunotherapy, Adoptive D009369:Neoplasms D000076962:Receptors, Chimeric Antigen D013601:T-Lymphocytes
nuclease homologous recombination CRISPR/Cas9 adeno-associated virus chimeric antigen receptor-T cells transcription activator like effector nuclease zinc-finger