Additional file 4: Fig. S4. Transformation efficiency of A. aculeatus TBRC 277 in the presence or absence of Cpf1 endonuclease containing plasmids. To investigate the toxicity of Cpf1 on the A. aculeatus host, 10-μg DNA of each plasmids were independently transformed into TBRC 277 protoplast. The number of transformants were recovered from minimal medium (MM+Czapek-Dox+bleomycin+sorbitol) supplemented with Uri/Ura. Protoplast transformed with empty vector, pCRISPR01, has no FnCpf1 gene (dark grey); protoplast transformed with FnCpf1-containing plasmid, pCRISPR01-FnCpf1 (light-grey); protoplast transformed with FnCpf1 and crRNA-pyrGs, pCRISPR01-FnCpf1-pyrGs (pyrG-1, pyrG-2, or pyrG-3) (white). The graph shows the means and standard deviation (SD) from two independent experiments.