Supplementary Figure 2.
Screening of mutations generated by the CRISPR/spCas9 system in the β-globin LCR HS2. (A) The locations of primer sets for PCR are presented on DNA with the targets equences of sgRNA and PAMs as described in Figure2. (B) Genomic DNA from clones was amplified by qPCR using the three primer sets, A1, A2 and A3. PCR product amounts were compared using IVR amplicon as an internal control and the nnormalized versus amounts in wild type (Wt) MEL/ch11 cells. (C) DNA sequences in the LCR HS2 are presented for wild type (Wt) cells and clones with GATA-1 or KLF1 motif deletions. The binding motifs of transcription factors are marked by colored bases. Black dashes are deleted nucleotides. (D) Genomic DNA of clones was amplified and quantified as described above. PCR products amplified by A4F/R primers were visualized in an agarose gel.