The first efficient non-lentiviral transduction system for immortal T-cells and B-cells.
No published method exists at present to efficiently transduce human lymphocytes with viral vectors containing DNA cargos large enough to deliver drug discovery assays. This is critical for preclinical studies in immuno-oncology (TCR, CAR-T) and human microbiome driven diseases revolving around inflammation. The data presented in the image below shows to our knowledge the first such method in immortal T-cells and B-cells.
Above Image: flow cytometry results for pseudotyped viruses with large DNA cargos transduced into T-cells (Jurkats). General transduction protocol (2.5 : 1 MOI). In a) the cluster corresponding to live cells was identified (orange), and only the events falling in this region were kept for analysis (b) 79.01% of the cells were transduced with a large DNA cargo (c-d): General transduction protocol with centrifugation (1000 rpm for 45 minutes, 20:1 MOI). 89.58% of the cells were transduced with a large DNA cargo. Centrifugation results in an increase in >10% transduction efficiency. (e-f): Negative control. The threshold for fluorescence was determined by setting the percentage of fluorescent cells in this sample close to 0.