Table 2.

CRISPR-Cas9 systems for genome editing in non-conventional yeasts.

Yeast	Cas9 expression	sgRNA expression	Gene disruption rate	HR rate	Reference
K. lactis	ScFBA1 promoter Genome integrated	SNR52 promoter	N/A	2% (3 integrations simultaneously) in NHEJ deficient strain	[35]
K. marxianus	ScTEF1 promoter codon-optimized	RPR1′-tRNAGly	66%	N/A	[62]
S. stipitis	eno1 promoter codon-optimized	SNR52 promoter	80%	N/A	[27]
Y. lipolytica	TEFintron promoter codon-optimized	TEFintron promoter, flanked by hammerhead and hepatitis delta virus ribozymes	85%	11% in wildtype up to 100% in NHEJ deficient strain	[60]
Y. lipolytica	UAS1B8-TEF promoter codon-optimized	SCR1′-tRNAGly promoter	92%	64% in wildtype up to 100% in NHEJ deficient strain	[61]
H. polymorpha	DH3 promoter human codon-optimized	tRNALeu	71%	47% (marker integration with selection)	[64]
P. pastoris	HTX1 promoter human codon-optimized	HTX1 promoter, flanked by hammerhead and hepatitis delta virus	100%	20%	[63]