Introduction: immunotherapy is a disease treatment methodology, based on activation or suppression of the host's immune system. Immune cell-based therapy is one of the most attractive immunotherapy strategies, because it can be tailored to target cancer cells specifically, thus avoiding any collateral damage associated with chemotherapy or radiation therapy. Development of induced pluripotent stem cell-based therapies has been the goal of many scientists since these cells were first obtained nearly twenty years ago. Induced pluripotent stem (iPS) cells can be derived via reprogramming of any somatic cell type and are capable of self-renewal as well as differentiation into any cell type. Additionally, because these cells are generated from adult organism cells, ethical problems and cell source abundance issues become irrelevant. Chimeric antigen receptors (CARs) are synthetic receptors that combine an antigen-binding domain with intracellular signaling domains, enabling immune cells to specifically recognize and target cancer cells. Therefore, CAR-iPS cell lines could serve as an excellent CAR source with the potential to be differentiated into CAR-expressing immune effector cells.

Objective: to establish three genetically edited novel CAR-expressing iPS cell lines.

Methods: CAR-coding genes were constructed from gBlocks. CAR-coding genes were inserted into an acceptor plasmid using In-Fusion cloning. Genome editing was performed with CRISPR‑Cas9. Template plasmid, Cas9 and sgRNA were delivered via electroporation. Selection of positive clones was performed with puromycin. Successful knock-in confirmed with genotyping.

Results: initial materials and strategies for establishing three CAR-iPS cell lines have been developed. Early CAR-iPS cell line derivation attempts show encouraging progress and will require further validation in the future.

Conclusion: CAR-iPS cell lines have the potential to serve as a reliable CAR cell bank.

Keywords: induced pluripotent stem (iPS) cells, chimeric antigen receptors (CAR), immunotherapy, genome editing.