Multiplex CRISPR/Cas9 gene-editing platform in oil palm targeting mutations in EgFAD2 and EgPAT genes

Journal of Genetic Engineering and Biotechnology

Table 2 The mutation rate of the four target sites in transformed oil palm tissues by biolistic, PEG, and Agrobacterium-mediated transformations analyzed using PCR and Sanger sequencing

Transformation method	Target gene	Constructs	No. of examined samples	No. of mutated samples	Mutation rate (%)	Mutants
Agrobacterium	EgFAD2	pYLCRISPR/Cas9PUbi-H: EgFAD2	18	5	28	2 (Double sgRNAs EgFAD2_T1T2) 3 (Single sgRNA_EgFAD2_T2)
	EgPAT	pYLCRISPR/Cas9PUbi-H: EgPAT	18	5	28	5 (Single sgRNA_EgPAT_T2)
	EgFAD2EgPAT	pYLCRISPR/Cas9PUbi-H: EgFAD2EgPAT	18	2	11	2 (Multigene: EgFAD2_T2, EgPAT_T2)
	Total		54
PEG	EgFAD2	pYLCRISPR/Cas9P35S-H: EgFAD2	6	2	33	2 (Double sgRNAs_EgFAD2_T1T2)
	EgPAT	pYLCRISPR/Cas9PUbi-H: EgPAT	7	0	0	-
	EgFAD2EgPAT	pYLCRISPR/Cas9PUbi-H: EgFAD2EgPAT	5	1	20	1 (Single EgFAD2_T1)
	Total		18
Biolistic	EgFAD2	pYLCRISPR/Cas9P35S-H: EgFAD2	48	0	0	-
	EgPAT	pYLCRISPR/Cas9PUbi-H: EgPAT	48	3	6	3 (Single sgRNA_EgPAT_T1)
	EgFAD2EgPAT	pYLCRISPR/Cas9PUbi-H: EgFAD2EgPAT	48	0	0	-
	Total		144

Mutation rate (%) = number of samples with mutations/number of total transgenic samples. Mutations were analysed by ICE