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Fig. 4 | Genome Biology

Fig. 4

From: Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease

Fig. 4

Germline transmission of Cas9 induced mutations from T1 to T2 plants in barley and B. oleracea in the absence of the T-DNA construct. a Sequence alignment of T1-181_E1 and five homozygous T2 progeny with homozygous 1-bp deletion in HvPM19_1. b Sequence alignment from representative clones of T1 heterozygote mutants and direct Sanger sequencing of their T2 progeny with homozygous mutations in the absence of the T-DNA. The numbers of clones supporting T1 mutant alleles are indicated on the right. c Sequence alignments of BolC.GA4.a Target 2 in homozygous T1 and T-DNA free T2 plants. Across panels the target sequences for sgRNAHvPM19-1 and sgRNABolC.GA4.a (grey) and PAM (red) are highlighted and Cas9 induced insertions and deletions are indicated by red font or red hyphens, respectively. Names of homozygous T2 plants that lack the presence of the T-DNA construct are indicated in blue; individual homozygous plants with the same allele are shown on the same row and are labelled with a ‘p’ prefix

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