Homology-dependent recombination of large synthetic pathways into E. coli genome via λ-Red and CRISPR/Cas9 dependent selection methodology

Table 1 Methods for the integration of large DNA module into E. coli chromosome

Method	Technique feature	Integration efficiency (%)	Integration sites	Maximum integration size (kb)	Rounds of integration	Marker left or markerless	Reference
I-SceI endonuclease	Combined λ-Red and I-SceI Cleavage	19.2–100	Desired locus	7	One	Markerless	[5]
λ-Red recombination	λ-Red assisted	0–50	Desired locus	7.3	One	Markerless	[7]
λ-Red recombination	Combined λ-Red and CRISPR/Cas9	10	Desired locus	7	One	Markerless	[33] [9]
		60		15.4	Five	Markerless
λ-Red recombination	λ-Red assisted	Not provide	fliK	15	Four	Markerless
		Not provide	fliK	50	Seven	Markerless
CRISPR/Cas9	Combined λ-Red and CRISPR/Cas9	60	Desired locus	7	One	Markerless	[12]
CRISPR/Cas9	Combined λ-Red and CRISPR/Cas9	50	Desired locus	10	One	Markerless	[13]
RecA homologous recombination	Combined λ-Red, RecA and CRISPR/Cas9	100	IS5	12	One	Markerless	This study

ISSN: 1475-2859