Skip to main content
. 2014 Jun 23;4:5400. doi: 10.1038/srep05400

Figure 4. Large deletions mediated by all-in-one CRISPR/Cas9 vectors.

Figure 4

(A) Schematic illustration of large deletion at the HPRT1 locus. The black boxes indicate exons. The blue lines and letters indicate PCR products. The zigzag lines indicate the HPRT1_A and HPRT1_B target sites. (B) Schematic illustration of large deletion at the APC locus. The black boxes indicate exons. The blue lines and letters indicate PCR products. The triangles indicate the off-5, off-8 and off-4 target sites. (C) Genomic PCR analysis of the HPRT1 and APC loci. The products from untransfected control cells (C) and cells transfected with the CRISPR/Cas9-nuclease vector expressing seven gRNAs (7) or the CRISPR/Cas9-nickase vector expressing six gRNAs (6) were analyzed by agarose gel electrophoresis. Red and blue asterisks indicate PCR products from un-deleted alleles, whereas yellow and green asterisks indicate PCR products from chromosomally deleted alleles. λ, λHindIII marker. W, Wide-Range DNA Ladder (100–2,000 bp) (Takara Bio). (D) Sequences of the PCR products from deleted alleles at the HPRT1 locus. The HPRT1_A and HPRT1_B target sites are indicated by red letters. PAM sites are indicated by black boxes. Deletions are indicated by dashes. (E) Sequences of the PCR products from deleted alleles at the APC locus. The off-5_L and off-4_R target sites are indicated by red letters. PAM sites are indicated by black boxes. Deletions are indicated by dashes.