Figure 4.

Multiplexing with a.Gag.MS2.CRISPR particles

(A–D) Efficient simultaneous knockout of two (B) or three (D) target genes by a.Gag.MS2.CRISPR particles. Prior to transduction of CXCR4⁺ Jurkat cells co-expressing RFP657.Tet2 and/or EGFP, the CRISPR RNA content of supernatants containing a.Gag.MS2 particles with two (A) or three (C) sgRNA.TS variants was determined. In total, knockout data of 4 to 5 individually packaged supernatants are depicted (n = 4–5). Representative flow cytometry plots of untransduced cells (mock) and of cells that were treated with the highest applied particle dose of the respective a.Gag.MS2.CRISPR supernatants are also shown. Errors bars in (B) indicate standard deviations.