CRISPR–Cas9-enabled genetic disruptions for understanding ethanol and ethyl acetate biosynthesis in Kluyveromyces marxianus

Table 2 KmADH and KmATF CRISPR–Cas9 target sequences and knockout efficiencies

Target	SgRNA	Gene disruption	Indel success
KmADH1	AGACTTCAAAGCCTTGTACA	WT CCGTGTACAAGGCTTTGAAG KO CCG--------GCTTTGAAG	2/10
KmADH2	GGTACCAGCTGGGATGTGAG	WT AAGCCGCTCACATCCCAGCT KO AAGCCGCT-----CCCAGCT	4/30
KmADH3	GCTATTCCAGAAAAGCAAAA	WT CAGAAAAGCAAAAGGGTGTT KO CAGAAAA-----AGGGTGTT	2/10
KmADH4	GCCATCCCAGAATCCCAAAA	WT TCCCAGAATCCC-AAAAGGG KO TCCCAGAATCCCAAAAAGGG	4/10
KmADH5	ATGGTCTTGAAAGAACACAA	WT GGTCTTGAAAGAA-CACAAG KO GGTCTTGAAAGAACCACAAG	1/10
KmADH6	GTACCACCACCGCAAAGTAG	WT CTACTTTGCGGTGGTGGTA KO CTACTT-GCGGTGGTGGTAC	2/10
KmADH7	GCTTGAGCTGAGAGATTGAT	WT AAGTCCCATCAA-TCTCTCA KO AAGTCCCATCAAATCTCTCA	3/5
KmATF	ATATAGTCTTCGGCAACACC	WT CGGTGTTGCCGAAGACTAT KO CGGTG--GCCGAAGACTATA	4/10

ISSN: 2731-3654