Table 3.

Genome editing at the endogenous CENH3 locus in hairy root lines of tomato

Line #	Deletions (bp)	In‐frame deletion present
1	∆7/∆7	No
2	∆12/∆12	Yes
3	∆7/∆6	Yes
4	∆12/∆12	Yes
5	∆152/∆152	Unknown
6	∆16/∆6	Yes
7	∆1/∆6	Yes
8	∆6/6	Yes
9	∆16/∆6	Yes
10	∆6/∆6	Yes
11	∆33/∆33	Yes

The guide RNA was designed to target the beginning of exon 4, at the beginning of the histone fold domain. Genomic DNA was extracted from independently derived hairy roots, and the endogenous CENH3 locus was amplified used primers flanking the gRNA target. PCR products that were not digested by the restriction enzyme BsmF1 (which recognizes the wt allele) were cloned into a plasmid vector and sequenced.