Fig. 1

Schematic diagram of gene repression via multiplex CRISPRi system in Y. lipolytica. The combination of four different repressors (dCpf1, dCas9, dCpf1-KRAB and dCas9-KRAB) and ten gRNAs that bind to different regions of gfp gene were employed in Y. lipolytica. But the results indicated that there was no clear correlation between the repression efficiency and targeting sites (left). As extra target sites screening often mandated a significant investment of time and effort, a strategy via one-step Golden-brick assembly of multiplex gRNAs was established for high-efficiency and tunable perturbation of gene expression in Y. lipolytica (right)