The resistance markers could ensure the entry of the CRISPR/Cas9 system into cells instead of gene editing. To increase the efficiency of positive colony screening on the primary transformation plates, we designed a visualized multigene editing system (VMS) via a unique tRNA-guide RNA (gRNA) array containing the gRNAs of a pigment gene and target genes. Disruption of produces white colonies, and the sequences of the endogenous tRNA, tRNA, tRNA, tRNA, and tRNA enhance gRNA release. The disruption efficiencies of multigene were analyzed in the strain AG11 using , , , , and as reporters. In white colonies on the primary transformation plates, the disruption rates of one-, two-, three-, four-, and five-target ... More
The resistance markers could ensure the entry of the CRISPR/Cas9 system into cells instead of gene editing. To increase the efficiency of positive colony screening on the primary transformation plates, we designed a visualized multigene editing system (VMS) via a unique tRNA-guide RNA (gRNA) array containing the gRNAs of a pigment gene and target genes. Disruption of produces white colonies, and the sequences of the endogenous tRNA, tRNA, tRNA, tRNA, and tRNA enhance gRNA release. The disruption efficiencies of multigene were analyzed in the strain AG11 using , , , , and as reporters. In white colonies on the primary transformation plates, the disruption rates of one-, two-, three-, four-, and five-target genes reached 89.2, 70.91, 50, 22.41, and 4.17%, respectively. The VMS developed here provides an effective method for screening homokaryotic multigene editing strains of .
