Softness limits CTLs self-killing and is inherited to T-leukemic cells for immune escape

Using CRISPR/Cas9-based gene overexpression method, we screened membrane or membrane-related protein molecule(s) by transfecting Jurkat cells with membrane protein gRNA library, followed by perforin treatment and PI staining. The top 10% among PI staining high and the bottom 10% among PI staining negative cells were sorted for next-generation sequencing (NGS) . Using the MAGeCK algorithm or RRA analysis, top sgRNAs were identified in PIh cells . We provide evidence that effector CTLs exhibit the perforin-resistant softness by downregulating cytoskeletal protein filamin A (FLNA)and this FLNA-dependent pathway is usurped by leukemic T cells to evade T cell killing. Using CRISPR/Cas9-based gene overexpression method, we screened membrane or membrane-related protein molecule(s) by transfecting Jurkat cells with membrane protein gRNA library, followed by perforin treatment and PI staining. The top 10% among PI staining high and the bottom 10% among PI staining negative cells were sorted for next-generation sequencing (NGS) . Using the MAGeCK algorithm or RRA analysis, top sgRNAs were identified in PIh cells . We provide evidence that effector CTLs exhibit the perforin-resistant softness by downregulating cytoskeletal protein filamin A (FLNA)and this FLNA-dependent pathway is usurped by leukemic T cells to evade T cell killing.