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. 2022 Jan 20;3:795644. doi: 10.3389/fgeed.2021.795644

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Copyright © 2022 Carlsen, Johansen, Yang, Liu, Westberg, Kieu, Jørgensen, Lenman, Andreasson, Nielsen, Blennow and Petersen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Two DMR6-1 gRNAs confer specific indel patterns. Two different gRNAs targeted at exon 1 in DMR6-1 were transformed into potato protoplasts and editing scored using IDAA. Grey peaks mark the wild-type amplicon. g43 confers a prevalence for a 3 bp deletion (upper panel), which in exons do not lead to frameshift of the coding sequence. g45 confers a high incidence of 1 bp indels (lower panel). These patterns were evident in repeated experiments (only data from one experiment shown). gRNA and IDAA primer pairs used: g43 and DMR6 forward primer + DMR6 reverse primer 2; g45 and DMR6 Forward primer 2 + DMR6 Forward primer 4 (see Material and Methods).