Fig. 4
From: Development of a CRISPR/Cpf1 system for targeted gene disruption in Aspergillus aculeatus TBRC 277
An example of A. aculeatus TBRC 277 protoplasts transformed with CRISPR/Cpf1 plasmids. a pCRISPR01-FnCpf1-pyrGs (with crRNA specific targeting pyrG gene) on primary selective transformation medium containing 1.2 M sorbitol + 10 mM uri/ura + 50 μg/ml bleomycin, b a single colony selected from panel a, was then cultured on new minimal medium containing 10 mM uri/ura+ 50 μg/ml bleomycin, (c) secondary selection from panel b, re-cultured on minimal medium containing 10 mM uri/ura and 1.5 mg/ml 5-FOA. d pCRISPR01-FnCpf1 encoding only Cpf1 (without crRNA targeting pyrG), after growing on secondary selective medium containing 5-FOA, no colony was observed (control). Uri: uridine, Ura: uracil