Figure 2 | Scientific Reports

Figure 2

From: Development of a versatile and conventional technique for gene disruption in filamentous fungi based on CRISPR-Cas9 technology

Figure 2

Establishment of the Cas9 stable expression Nodulisporium sp. (No. 65-12-7-1). (a) Analysis of the Cas9-eGFP expression in the mycelia transfected with pDHt/sk-Ppdc-toCas9-eGFP-Tpdc (i, ii) and wild-type (iii, iv). The bar represents 100 μm. (b) Analysis of the integration of cas9 gene into the genome. (lane 1: wild-type; lane 2: pBSKII-tocas9-hph; lane 3: JN1002; lane 4: JN1001; lane 5: JN1003; lane 6: JN1004; lane 7: JN1005; lane 8: JN1006; lane 9: JN1007). (c) RT-PCR analysis of the Cas9 expression. Upper panel for GAPDH and lower panel for Cas9. lane 1: H2O; lane 2: pBSKII-toCas9-hph; lane 3: genomic DNA of wild-type; lane 4: genomic DNA of JN1001; lane 5: RNA of wild-type; lane 6: RNA of JN1001; lane 7: cDNA of wild-type; lane 8: cDNA of JN1001. (d) Real-time PCR quantitative analysis of Cas9 expression in WT and JN1001.

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