Fig. 1

High temperature affects genome editing efficiency. A Heat shock protocol. Every line is either subjected to a series of three heat shocks (indicated with red squares) or is grown under normal conditions (21°C) in a Tissue Culture (TC) room. Each 24h heat shock at 37°C in a bacterial incubator is alternated with a 24h recovery period in a TC room (21°C). After the final heat shock, plants are allowed to recover and grow for ~14 days, followed by visual scoring of phenotypes (if possible) and/or genotyping by Sanger sequencing. B Effect of number of consecutive heat shocks on genome editing efficiency. Segregating Arabidopsis T2 lines expressing PcUBI::LbCas12a::G7T (four independent lines, Cas12a #1-#4) or PcUBI::Cas9::G7T and a gRNA targeting PDS3 were grown under control conditions (21°C) or subjected to one, two, three or four heat shocks (1-4xHS; 37°C). After 14 days of recovery, plants displaying a pds phenotype were scored. n=75 per line per treatment (bars). DNA was extracted for eight randomly selected individuals for each line and treatment and PCR products amplified from targeted loci were sequenced and analysed using ICE (https://ice.synthego.com). The KO-score is given for each sample (dots), which indicates those indels that result in a frameshift or are 21+bp in length. n=8 per sample per treatment