GoKawiila, Different promoters used to evaluate the toxicity and the immune activity of CRISIS systems. The source organism of each tested CRISIS system is listed. b, Transformation efficiency of E. coli cells transformed with a pET28a derivative over-expressing various CRISIS genes. Data are presented as mean ± s.d. from three biological replicates. c, Serial dilutions were plated on non-selective agar plates to assess growth impairment caused by IPTG-induced CRISIS systems. The experimental design is depicted on the top. Cultures of transformants obtained from Kan+/IPTG− plates were plated on non-selective agar plate with or without IPTG to assess cell survival. The strain harboring the empty vector served as the negative control. d, Measuring NAD+ levels in E. coli cells overexpressing Sir2-HerA versus the empty vector control. Data are presented as mean ± s.d. from three biological replicates. The P value was calculated using a two-sided Student’s t test. e, Measuring NAD+ levels in E. coli cells harboring pCas-Sir2-HerA after a 3-hour induction of AcrIC9 (from another plasmid) using varying IPTG concentrations. Data are presented as mean ± s.d. from three biological replicates. P values were calculated relative to the acr- control using a two-sided Student’s t test. f, Plating of Wukong-expressing cells on selective and non-selective plates, with or without IPTG.
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