Fig. 4: Signal joint formation and isolation using an integrated target. | Nature Communications

Fig. 4: Signal joint formation and isolation using an integrated target.

From: Generating combinatorial diversity via engineered V(D)J-like recombination in Saccharomyces cerevisiae

Fig. 4

a Diagram of the integrated SJUE target in chromosome XV (Chr XV). Cutting by RAG1/2 can lead to NHEJ repair which fuses the RSSs together and forms a signal joint. Such an event permanently removes eGFP and URA3, leading to cells that do not fluoresce and are 5-FOA-resistant. b The SJUE target was integrated in both a recombination-competent (AC518: NLS-RAG1core, RAG2, and HMGB1) and recombination-incompetent (BY4742) strain. After an 8-d induction in YPG media, cells were plated on SD, 5-FOA plates. Both eGFP-positive and eGFP-negative colonies were separately counted. CFU = colony forming unit. In (b), data are presented as mean values ± SD; n = 3 biological replicates. Statistical significance was calculated using a two-way ANOVA with repeated measures across fluorescence types and Fisher LSD test. Source data are provided in the Source Data file.

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