Figure 3 | Scientific Reports

Figure 3

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

Figure 3

Efficient gene disruption by co-transformation of in vitro transcriptional gRNA and the linear marker gene cassette. (a) Analysis of the integration of neo gene into the genome of G418-resistance clones generated by co-transformation of the in vitro transcriptional gRNA and linear neo cassette into the JN1001. (b) PCR amplification of the DNA regions surrounding the target site of the clones described in (a) using primers flanking the target site. (c) Analysis of the integration of neo cassette into the genome of G418-resistance clones generated by co-transformation of the in vitro transcriptional gRNA and circular plasmid pBSKII-PtrPC-neo-TtrPC containing the neo cassette into the JN1001. (d) PCR amplification of the DNA regions surrounding the target site of the clones described in (c) using primers flanking the target site. (e) Sequence analysis of PCR products generated in (b). (f) Analysis of the effects of usage amount of linear neo cassette on the mutation efficiency. ((a–d), lane 1: JN1001; lane 2-13: G418-resistance clones (No. 1-12)).

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