Figure 1.

Overview of experimental setup. Cells are transfected with (A) guide RNA expression vector(s) under the control of hU6 promoter, (B) a Cas9-ginβ expression vector under the control of a CMV promoter and (C) a recCas9 reporter plasmid. Cotransfection of these components should result in (D) reassembly of guide RNA-programmed recCas9 at the target sites. This will mediate deletion of the polyA terminator, allowing transcription of EGFP. Guide RNA expression vectors and guide RNA sequences are abbreviated as gRNA.