Figure 5
Prompt characterization of TCSs in E. coli with the ddCpf1-mediated multiplex gene repression strategy. The 32 RRs in E. coli were divided into 6 groups (Supplementary Tables S3 and S4), and the growth phenotypes were analyzed by culture in either rich LB medium (a) or minimal M9 medium (b). In c, cells expressing ddCpf1 and individual crRNAs that targeted genes in arrayG5 were cultured in M9 medium, employing arrayG5 as a control. The growth-deficient strains, which expressed ddCpf1 and arrayG5 (or crRNAs targeting genes in arrayG5), were marked with the red dashed box, while the yellow dashed box marked the slow growth phenotype of arrayG6. A gradient dilution of cells with sterile water was performed, and different amounts of cells (that is, 4.0×105, 4.0×104, 4.0×103, 4.0×102 and 4.0×101) were dotted on plates. Cells expressing ddCpf1 only were employed as the positive control.
