Metabolic engineering of Komagataella phaffii for enhanced 3-hydroxypropionic acid (3-HP) production from methanol

Table 2 Variable sgRNA targets custom-designed in this study

Target name	Locus	Sequence (5’ ◊ 3’)	Module inserted
pENO1^UP_sgRNA1	PP7435_Chr3-1154	TTATTGAAGATACCGCCCGG	P_DAS1-AAT2-RPS3tt P_DAS1−AspDH_Spe-RPS3tt
pENO1^UP_sgRNA2	PP7435_Chr3-1154	CGTTTAAACTTACCTCCGGG	P_DAS2-JEN1- ScCYC1tt P_DAS1-ESBP6_Sc- ScCYC1tt
AOX1tt_DOWN_sgRNA1	PP7435_Chr4-0131	ACTATTGATCCAAGCCAGTG	P_SHB17-PYC2-RPS3tt
AOX1tt_DOWN_sgRNA2	PP7435_Chr4-0131	GCTGATTTGGGGTTTAATAC	P_SHB17-PYC2-RPS3tt
GUT1tt_DOWN_sgRNA1	PP7435_Chr4-0174	GGTAGATGAGTTATTAACTG	P_DAS2-JEN1-ScCYC1tt P_DAS1-ESBP6_Sc-ScCYC1tt
GUT1tt_DOWN_sgRNA2	PP7435_Chr4-0174	AAACTGCGAATAAGAAAGCA	P_DAS2-JEN1-ScCYC1tt P_DAS1-ESBP6_Sc-ScCYC1tt
pOPT1-1^UP_sgRNA1	PP7435_Chr4-1011	ACTAACCAGAGCAAACACTG	P_DAS2-JEN1-ScCYC1tt
pAOX1^UP_sgRNA1	PP7435_Chr4-0130	ATTGTGAAATAGACGCAGAT	P_FDH1-FDH(V9)_Pse-TDH3tt
pAOX1^UP_sgRNA2	PP7435_Chr4-0130	GCAGTCGATCTCAAAAGCAA	P_FDH1-FDH(V9)_Pse-TDH3tt
ΔFDH1_sgRNA1	PP7435_Chr3-0238	ATCGGCGGCGTGCTTACCAG	-
ΔFDH1_sgRNA2	PP7435_Chr3-0238	GTTCTCGTTTTGTACTCCGC	-

ISSN: 1754-1611