Targeted transposon screening in bacteria by CRISPR/Cas12k-guided transposase

Here, we report a CRISPR/Cas12k-transposon-assisted genome engineering (CTAGE) method that allows for high-throughput site-specific mutagenesis in microbial genomes. Exploiting the powerful CTAGE technique, we construct a site-specific transposon mutant library focusing on all the possible transcription factors (TFs) in Pseudomonas aeruginosa, enabling comprehensive identification of essential genes and new factors for antibiotic resistance. Overall design: Examination resistance gene of imipenem, ceftazidime and ofloxacin in Pseudomonas aeruginosa.