Figure 5.

dCas9-Spo11 fusion binding at various genomic loci. Quantitative analysis of dCas9-Spo11 association with the target GAL2 promoter upon expression of the gRNA handle or multiple gRNAs (A), and with various genomic loci upon expression of the corresponding gRNAs (B), was assessed by Chromatin immunoprecipitation (ChIP) in sae2Δ diploids expressing the Flag-tagged dCas9-Spo11 protein. Cells were collected at the indicated times after transfer to sporulation medium (t = 0 h), fixed with formaldehyde in vivo and immunoprecipitated with anti-flag antibodies. Quantitation of immunoprecipitated DNA was performed via qPCR and fold enrichments are expressed as the relative fold enrichment of target site over SMC1 signal and are normalized to input DNA (Materials and Methods). In the panel B the enrichment for each target site was normalized to the fold enrichment measured in control gRNA handle-dCAS9-SPO11 cells.