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Additional file 7: Table S3. The sequence of FnCpf1, eGFP, and pCRISPR01 expression vector.

Additional file 1: Fig. S1. Schematic representation of pCRISPR01 plasmid construction backbone for CRISPR/Cpf1 genome editing experiment in A. aculeatus TBRC 277.

Additional file 5: Table S1. CRISPR/Cpf1 gene editing efficiency targeting pyrGof A. aculeatus TBRC 277.

Additional file 3: Fig. S3. The functional plasmid construction of CRISPR/Cpf1 system. The plasmid consists of Cpf1 nuclease gene and crRNA-pyrG cassettes for targeted genome modification in A. aculeatus TBRC 277. (a) pCRISPR/Cpf1-pyrG plasmid, crRNA is expressed under control of U3-AF pol. III promoter, (b) Guide RNA cassette, crRNA, consists of 19-bp direct repeats (DR) to form a stem-loop structure, as well as 20-bp pyrG-targeted protospacers.

Additional file 6: Table S2. List of primers used in this study.

Additional file 2: Fig. S2. Western blot detection of heterologous EGFP and Cpf1-EGFP proteins in recombinant A. aculeatus TBRC 277 egfp and Cpf1-egfp, respectively. (a) GenCRISPRTM FnCpf1 monoclonal antibody (9H6) was used as the primary detection, 1.0 μg (Genscript, USA). (b) GFP monoclonal antibody (C163) was used for the primary detection, 3 μg (Thermo Fisher, USA). All samples were treated by IgG-AP as the secondary antibody for detection, 2 μg. Lane 1, crude protein of A. aculeatus recombinant (Cpf1-egfp). Lane 2, crude protein of recombinant A. aculeatus (egfp), Lane 3, crude protein of A. aculeatus TBRC 277 wild-type (control). Each lane was loaded with 20-μg protein. Anti-GFP (Roche) and FnCpf1 (Genscript, USA) antibodies were used as the primary (monoclonal) antibodies, anti IgG-conjugated AP was used for secondary antibody. M: PageRule Plus Prestained Protein Ladder (Thermo Fisher, USA).