Three major insights: 1) CAST-based barcoding and gene disruption is straightforward, but requires screening and groundwork for quantitative use. 2) The Tn7 insertion locus, when it exists, needs critical evaluation. 3) “tagIMseq” is a versatile, rapid, and easy transposon mapping method
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![A hyperactive ACD6 allele reduces growth and causes necrosis only in the greenhouse. Top: Representative Arabidopsis thaliana rosettes from different genetic backgrounds. Yellow and blue labels represent descendants of a heterozygous ACD6 HIF line that are homozygous for either the Est-1 or the Col-0 allele of ACD6. Green and purple labels represent the Est-1 accession and the isogenic acd6 knockout null allele in that accession. Plants were grown in Tübingen in the greenhouse (GH-Tü), outdoors in the field in Tübingen (F-Tü), or outdoors in the field in Zurich (F-Z, a GMO-approved facility). Note the necrotic older leaves for greenhouse plants with the ACD6-Est-1 allele (red arrows). Bottom: Rosette size of individual plants, calculated by quantifying green pixels from images similar to those in the top panel. The line within each box represents the median. Boxes enclose the interquartile range (IQR) with whiskers extending to up to 1.5 times the IQR. Brackets indicate Mann–Whitney U-test results, with * indicating P < 0.05 after Benjamini–Hochberg correction for multiple testing across the four comparisons. Dotted lines extending from the mean of each distribution span the observed effect size, and a bootstrapped distribution of computed effect sizes [30] is shown in gray, centered on the mean of the second box in each pair (red dot). The black bar extending vertically from the red dot represents the 95% confidence interval of the effect size.](https://cdn.bsky.app/img/feed_thumbnail/plain/did:plc:5522ztebtekoor5efelihqhb/bafkreiaqunadge3nltwergaw3byzldjmd7k2prjfsz7tfv4ycecjvwxmtm)
