Pooled CRISPR Interference Screening Identifies Crucial Transcription Factors in Gas-Fermenting Clostridium ljungdahlii

NGS profiling of crRNA library (Figure S1); flowchart illustrating the screening process for identifying the TF genes associated with the heterotrophic growth of C. ljungdahlii (Figure S2); influence of the repression of six selected TF genes on the heterotrophic growth of C. ljungdahlii in the YTF medium (Figure S3); flowchart illustrating the screening process for identifying the TF genes associated with autotrophic growth of C. ljungdahlii in gas fermentation (Figure S4); influence of the repression of 20 selected TF genes on the growth rate of C. ljungdahlii in gas fermentation (Figure S5); influence of the overexpression of 13 selected TF genes on the product (acetate and ethanol) formation of C. ljungdahlii in syngas fermentation (Figure S6); and the scoring matrix used to measure the significance of off-target hits (Figure S7); A CRISPRi library designed to target transcription factors (TFs) in C. ljungdahlii encompasses a total of 4153 crRNAs, which is directed at 425 TF genes (Table S1); CRISPRi library designed to target 400 negative control crRNAs with no binding hits in the C. ljungdahlii genome (Table S2); 75 TF genes have potential associations with the heterotrophic growth of C. ljungdahlii (Table S3); 68 TF genes have potential associations with the autotrophic growth of C. ljungdahlii on gas fermentation (Table S4); 33 TF genes play crucial regulatory roles in C. ljungdahlii grown on both sugars and C1 gases (Table S5); 35 TF genes appear to be specifically associated with the autotrophic growth of C. ljungdahlii in gas fermentation (Table S6); knockdown of 18 TF genes through CRISPRi significantly altered cell growth rate of C. ljungdahlii in gas fermentation (Table S7); strains and plasmids used in this study (Table S8); and primers used in this study (Table S9) (PDF)