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. Examination resistance gene of imipenem, ceftazidime and ofloxacin in Pseudomonas aeruginosa.