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. 2022 Dec 22;9(1):16. doi: 10.3390/jof9010016

Figure 4.

Figure 4

Mad7-induced gene editing of the albA locus in A. niger. (A) Strategies for introducing a specific XbaI site/amber stop codon into albA (left) and an mRFP insertion (right) into albA locus. A DNA DSB is introduced in albA locus by Mad7 containing an albA specific gRNA. A single-stranded oligonucleotide is used as RT to direct introduction of the XbaI site/amber stop codon by HR. A PCR fragment is used as RT to direct insertion of mRFP into albA by HR. See main text for further details. (B) Mutation of albA changes the conidia color of transformants growing on solid medium from wild-type black to white. (C) To the left, transformation controls with empty Mad7-CRISPR plasmid (pDIV300) imaged in visible and red fluorescent light as indicated; and with the albA-Mad7-CRISPR plasmid (pDIV313) in the absence of a (RT). In the middle, introduction of a stop-codon mutation into albA using the albA Mad7-CRISPR plasmid (pDIV313) and an oligonucleotide (PR_DIV3196)-based RT. To the right, insertion of mRFP into albA using the albA-Mad7-CRISPR plasmid (pDIV313) and the PCR-based RT. Plates were imaged with visible and red fluorescent light as indicated.