Fig. 1
From: A modular toolbox for gRNA–Cas9 genome engineering in plants based on the GoldenBraid standard
Multiple guide RNAs assembly with GoldenBraid. a Software-assisted CRISPR cloning workflow. Targets are adapted to the GoldenBraid standard with the ‘GB-CRISPR domesticator’. Then, these level 0 parts (D/M-Targets) are combined with other standard GBparts with the ‘GB-CRISPR assembler’ to create the guide RNA expression cassettes, which can be combined between them and/or with a Cas9 transcriptional unit with the ‘GB-binary assembler’. b Optimization of GB-CRISPR multipartite reactions. Forward and reverse primers were diluted to different concentrations with different solvents; they were mixed and twelve independent multipartite reactions were set up. After transformation into E. coli, the number of colonies was estimated. c Number of colonies obtained on the twelve independent guide RNA multipartite assembly reactions