Figure 4.

Evaluation of Eight gRNAs in Various Polycistronic Expression Systems and the Effect of Position in the Array on gRNA Activity.

(A) Structure of seven constructs for expressing gRNAs 49 to 56 targeting four tomato genes for deletion.

(B) Detection of deletions between gRNA sites using qPCR. Each gene is targeted for deletion with two gRNAs designed to create a 3-kb deletion to prevent amplification of the nondeleted, wild-type template. Deletion frequency was quantified by qPCR using primers shown as red and blue arrowheads.

(C) Deletion frequencies in tomato protoplasts as measured by qPCR. Positions of respective gRNAs in the array are specified for each construct. Error bars represent se of two replicates.