Figure 1.

Cas12a genome editing system. (A) FnCas12a and crRNA were expressed from a single plasmid with ura4 marker and ars1 replicating origin in fission yeast. As the first construct, FnCas12a and gRNA were expressed with adh1 and rrk1 promoter, respectively. (B) To test editing efficiency, we picked four gRNAs targeting the endogenous ade6⁺ CDS. One linear donor DNA was co-transformed to delete ade6 by homologous recombination. (C) The gRNA sequences used and their editing efficiency, with mean values ± S.E.M. The original colony counts are listed in Supplementary Table S6. (D) Colony PCR to check the ade6 deletion, in red colonies and white colonies from plates with PMG5–Ura w/ low adenine media. The 1 kb Plus DNA ladder from NEB (Catalog# N3200L) was used as the molecular weight standard. (E) Spot assay on plates to check adenine auxotroph.