Table 1. Efficiency obtained before and after selection of 5-FOA by PCR amplification at the mutated locus and sequence verified.

Gene targeting	Constructs	sgRNA	Method	Before 5-FOA	After 5-FOA
albA− Codon stop insertion	cDNA006 X-pyrG-X	sgRNA001	Non-selective	19 white colonies 5 black colonies (79% white colonies)	8 white colonies 2 black colonies
albA− Deletion of 1000 bp	cDNA008 X-pyrG	sgRNA001	Selective	20 white colonies 8 black colonies (71% white colonies)	7 white colonies 0 black colonies
ΔglaA Gene replacement with A5IL97	cDNA009 A5IL97-X-pyrG	sgRNA002 sgRNA003	Selective	8 colonies with pyrG at the locus 2 colonies without pyrG at the locus	8 on target 0 off target
ΔmstC Gene deletion	cDNA010 X-pyrG	sgRNA004 sgRNA005	Selective	7 colonies with pyrG at the locus 3 colonies without pyrG at the locus	7 on target 0 off target