FIGURE 1.
Schematic of CRISPR-Cas12a-assisted genome editing in A. mediterranei U32. First, FnCas12a was integrated into U32 chromosome and constitutively expressed under the promoter of Papr. Then, the crRNA transcribing plasmid was electroporated into the U32 competent cells that harbored FnCas12a. Guided by a target-specific crRNA, Cas12a specifically cleaved target dsDNA and generated DSBs on the chromosome, which could be repaired by either NHEJ or HDR, generating desired mutants. When DSBs were repaired by NHEJ, the breaks were directly jointed with no homologous donor DNA required; however, DNA sequences of ranging length around the Cas12a cleavage site will be deleted during the repair.