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. 2019 Jan 1;30(1):119–130. doi: 10.1091/mbc.E18-09-0605

FIGURE 2:

FIGURE 2:

CRISPRi-mediated knockdown of exogenously expressed NanoLuc in Giardia. Nine gRNAs were designed to repress transcription of the luciferase reporter gene NanoLuc expressed exogenously from the Giardia MDH promoter (A). Eight gRNAs targeted the coding region of the NanoLuc gene and one gRNA (-11) targeted the region immediately upstream of the translation start site. Target positions are numbered relative to the translational start site ATG and are located three bases upstream of the PAM sequence (NGG) for each gRNA. Relative luminescence per cell is plotted for each gRNA designed to repress Nanoluc expression. In B, the mean luminescence per cell for two independent transformations is compared between the different luciferase knockdown strains. Error bars indicate 95% confidence intervals. The significance of luciferase knockdown relative to a nonspecific gRNA control was assessed using an unpaired t test, with ns = not significant, *p ≤ 0.05, **p ≤ 0.01 (Materials and Methods).