Fig. 2
From: Engineering Halomonas species TD01 for enhanced polyhydroxyalkanoates synthesis via CRISPRi
CRISPRi system used for Halomonas sp. TD01 (a) and relative binding positions of sgRNAs targeting ftsZ gene (b). Pli-dCas9-sgRNA, plasmid carrying the CRISPRi system; Ptrc, trc promoter; CmR, chloramphenicol resistance gene; oriT, origin of transfer; ftsZ, filamenting temperature-sensitive mutant Z. The length of ftsZ gene is 1179 bp, while the length of sgRNAs is around 20 bp. The promoter of ftsZ gene is 35 bp to 10 bp upstream of ftsZ gene. To inhibit ftsZ gene expression, ftsZ1 is designed 4 bp upstream from ATG sequence, from position −26 to −5, after the PAM sequence CGG (from position −29 to −27). FtsZ2 is designed 89 bp downstream of ATG sequence, from position 90 to 112, after the PAM sequence TGG (from position 87 to 89)