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CRISPR/Cas9 coupled λ-Red recombination technology had been utilized to execute genome editing in various genetics. For finding a competent solution to modify the virulence genetics of enterotoxigenic E. coli (ETEC), the two-plasmid system was utilized. The coding series (CDS) area for the estA, eltI, estB, eltIIc1, and faeG locus were deleted. The coding area of estB ended up being substituted with estA. Gene recombination performance ranged from 0 to 77.78% as soon as the duration of the