FIGURE 2.

The CRISPRi screening procedure in bacteria. Firstly, oligos designed on a large scale by the software are synthesized through a chip and then, the sgRNA plasmids library is constructed. Mix sgRNA plasmids are co-transformed into recipient strains in which dCas9 is integrated into genomic DNA. Competitive growth culture is performed in selective condition and control conditions. The mixed plasmids are isolated from pooled colonized strain and subsequently used for high-throughput sequencing. Finally, data analysis of the relative fitness contribution of each gene is performed.