Link to open-access version of our paper: rdcu.be/eHZHx

This is our proudest work yet—a blueprint for rationally designing next-generation cell therapies. Data and methods are fully open-source for academic use and licensable for translation. Out in @nature.com today. www.nature.com/articles/s41...

Great teamwork: P Datlinger*✉️, E Pankevich*, C Arnold*, N Pranckevicius, J Lin, D Romanovskaia, M Schäfer, F Piras, AC Orts, A Nemc, P Biesaga, M Chan, T Neuwirth, A Artemov, W Li, S Ladstätter, T Krausgruber & C Bock✉️ (*first authors, ✉️correspondance).

CELLFIE supports diverse CRISPR modalities. Base editing proved especially powerful. Tiling screens of RHOG suggest inhibition of GTP binding pocket as therapeutic angle and identified gRNAs for clinical translation. (7/7)

With many promising single edits reported across labs, the key challenge is combining them into tailored therapeutic designs. We extended CELLFIE with combinatorial screens and discovered a strong synergy between RHOG and FAS knockouts. (6/7)

RHOG was a surprising hit and has been linked to immunodeficiency in humans. Yet in CAR T cells, RHOG knockout drives proliferation, fosters a central memory phenotype, and reduces exhaustion. (5/7)

We developed in vivo CROP-seq for screens in mouse immunotherapy models. By reading gRNAs from abundant mRNA and tracking individual clones, we identified RHOG, FAS, and PRDM1 as CAR T boosters. In vivo CROP-seq supports discovery screens with thousands of perturbations. (4/7)

To address clinical limitations of CAR T therapies, we performed genome-scale FACS screens for target recognition, T cell activation, apoptosis/fratricide, and exhaustion—resulting in the largest CAR T screening dataset to date. (3/7)

We ran genome-wide screens for T cell fitness under TCR and repeated target cell stimulation. This revealed novel knockouts that enhance CAR T cell persistence, alongside many well-known clinical targets. (2/7)