Figure 6.
Multiplex KO was highly efficient by Cas9 RNP nucleofection.(A) Double KO of TIGIT and CD96 by combining two doses of 40 pmol Cas9 RNP complexes in one nucleofection reaction. Representative flow cytometry plots show the mean percentages of positive and negative cells. (B) Double KO of TIGIT and CD226 (encoding DNAM-1). (C) Triple KO of TIGIT, CD96, and CD226 by combining three doses of 40 pmol Cas9 RNP complexes in one nucleofection reaction. Q2 and Q4 of the representative flow cytometry plots were gated to determine triple-positive and triple-negative cells, respectively. Data are shown as mean ± SD of three donors (n = 3). Two-tailed Welch’s unequal variances t test was used to test for statistical significance. FSC, forward scatter; ns, not significant.

Multiplex KO was highly efficient by Cas9 RNP nucleofection. (A) Double KO of TIGIT and CD96 by combining two doses of 40 pmol Cas9 RNP complexes in one nucleofection reaction. Representative flow cytometry plots show the mean percentages of positive and negative cells. (B) Double KO of TIGIT and CD226 (encoding DNAM-1). (C) Triple KO of TIGIT, CD96, and CD226 by combining three doses of 40 pmol Cas9 RNP complexes in one nucleofection reaction. Q2 and Q4 of the representative flow cytometry plots were gated to determine triple-positive and triple-negative cells, respectively. Data are shown as mean ± SD of three donors (n = 3). Two-tailed Welch’s unequal variances t test was used to test for statistical significance. FSC, forward scatter; ns, not significant.

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