Figure 3 | Scientific Reports

Figure 3

From: A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus

Figure 3

Simultaneous double virulent gene recombination using CRISPR/Cas9 System.

(a) Procedure of CRISPR/Cas9 system assisted double-gene recombination. (b) HEK293T cells co-transfected with sgRNAs and DNA donors and then infected with PRV HNX (MOI = 1). Upon recombination, GFP and mCherry expression will be driven by viral TK and gE promoter respectively. (c) Single cells with fluorescent gene recombinant virus were sorted by FACS, plated to 96 well plate pre-cultured with PK15 cells and incubated until fluorescence appeared. (d) Plaque purification of double genes recombinant virus in agarose-DMEM plates. Cell plaques were indicated by arrows. (e) PCR verification of TK and gE gene deletion.

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