Figure 7
In vivo detection of pp38 mutation. Loss of the PspF1 restriction site was detected in cDNA constructed using RNA templates from PBMCs. PBMCs were collected from the ST-1st group, and total RNA was isolated and used to synthesize cDNA. The pp38 ORF was amplified by PCR using cDNA as the template. Positive samples were excised from gels, reamplified and digested with the restriction enzyme PspF1. Resistant DNA demarcated the loss of the restriction site due to ST-mediated CRISPR interference. The pp38 ORF from wild-type MDV was treated as the control.