Fig. 4. Deep-sequencing analysis of the PVX-SpCas9 mediated GE. (A) Frequency of sequence variations by length detected by amplicon deep sequencing of PDS-sgRNA1 target region in N. benthamiana leaves treated with heat stress (HS) and no heat stress (NHS). (B) Frequency of sequence variations by length detected by amplicon deep sequencing of PDS-sgRNA2 target region in N. benthamiana leaves treated with HS and NHS. (C) Percent GE recorded with NbPDS-sgRNA1 and NbPDS-sgRNA2 constructs. (D) Alignment of Mi-seq sequencing reads showing the presence of indels at the NbPDS-sgRNA1 target sequence. (E) Alignment of Mi-seq sequencing reads showing the presence of indels at the NbPDS-sgRNA2 target sequence. The number following the sequence variant and underscore indicates the total number of that particular variant. Target sequence with the PAM is shown in green and protospacer is marked with red font.
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