The editing of CD4+ T cells has shown high efficiency and stability, with a robust ability to maintain functionality after the editing process. The efficiency of the CD4+ T cell editing was determined through ICE analysis, with results demonstrating high editing efficacy across multiple loci while ensuring cell viability during the freezing stage. Post-editing, the cells can be thawed and expanded without loss of functionality or editing efficiency. Cytokine production in edited T cells, including IFNg, IL2, and TNFa, indicates that the editing process does not negatively impact the cells’ ability to produce essential cytokines upon stimulation. These edited pools also show high viability, making them suitable for further downstream assays.

• – High editing efficiency of CD4+ T cells determined by ICE analysis.
• – No reduction in cell viability after freezing, as indicated by flow cytometry
• – Successful expansion and thawing of edited cells without affecting functionality
• – Edited cells exhibit high levels of cytokine production (IFNg, IL2, TNFa) post-stimulation 
• – High viability and expansion potential, with >90% viability after 14 days in culture 
• – Statistical analysis shows significant cytokine differences between edited and mock cells