Extended Data Fig. 4: Benchmarking DART vectors.
From: Species- and site-specific genome editing in complex bacterial communities
a, E. coli WM3064 to E. coli BL21(DE3) conjugation, transposition, and selection schematic (top) and guide RNAs targeting the lacZ α-fragment of recipient BL21(DE3), which is absent from donor WM3064 (bottom). b,d,f, Percent selectable transposed colonies is calculated as the number of colonies obtained with gentamicin selection divided by total viable colonies in absence of selection. b, Insertion-receiving colonies divided into on- and off-targeted. This was calculated by multiplying % selectable colonies for representative guides in d and f (highlighted by grey bars) by the on- or off-target rates (shown in Fig. 2b). c, Transposition with VcDART was tested using three promoters. The variant using the Plac promoter, harvested from pHelper_ShCAST_sgRNA16, was also used for Figs. 2–5 and Extended Data Figs. 4b, 5, 6, and 8. d, Efficiencies of VcDART using various promoters. e, Transposition with ShDART was tested with three transcriptional configurations, all using Plac16. The configuration used for characterization of ShCasTn originally16 was also used for Fig. 2 and Extended Data Fig. 4b. f, Efficiencies of ShDART using various promoters. b, d, f, Crossbar indicates mean and error bars indicate one standard deviation from the mean (n = 3 biological replicates). Guide RNAs ending in ‘NT’ are non-targeting negative control samples.
