Figure 1.

Optimization of electroporation conditions

(A) Disruption of TRAC locus. Activated T cells were electroporated with TALEN-encoding mRNA (7.5 μg) using three different settings. Edited T cells were harvested at indicated time points and extent of T cell receptor knockout quantified by measuring CD3 expression. (B and C) Cell viability and cell proliferation. Electroporation-associated toxicity (B) and cell numbers (C) were determined using an automated cell counter (NucleoCounter). (D) Titration of the TRAC-targeting TALEN mRNA. Following electroporation with the indicated amounts of TALEN RNA (setting 3), cells were harvested and extent of T cell receptor knockout quantified by measuring CD3 expression. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001. UT, untreated T cells; Set.X, electroporation settings (see Table 1).