Gene stacking in plant cell using recombinases for gene integration and nucleases for marker gene deletion

BMC Biotechnology

Table 1 I-SceI and ZFN activity at the site-specific integration locus

Lines	Nuclease	Expression system¹	Total #	Perfect deletion²	Short indels³	Large indels⁴	No change⁵
UI	I-SceI	Constitutive	72	14	7	25	26
UZ	ZFN	Constitutive	68	4	5	38	21
HI	I-SceI	Heat-shock	7	3	-	-	4
HZ	ZFN	Heat-shock	23	-	-	13	10

¹Strong constitutive (Ubi promoter) or heat-shock (HS) expression
² < 15 bp indel at the target without the loss of sequences immediately adjacent to the nuclease target
³50–200 bp insertion or deletion
⁴No amplicons obtained in PCR
⁵No DSB-repair as indicated by PCR and/or sequencing

ISSN: 1472-6750