Fig. 1: Graphic summary of the mutagenesis system development.
a Schematic representation of the mutagenic process. The T7 RNA polymerase fusion (BD-T7RNAP, blue shape joined by a black line to a purple elliptical shape) specifically binds the T7 promoter (i), initiating the transcription and moving along the target gene (yellow filled arrow) carrying the base deaminase (BD) that introduces mutations (red stripes) in the gene (ii). The fusion stops and detaches from the DNA when encounters a dCas9 molecule (translucid ovoid shape) bound to a specific sequence determined by the CRISPR RNA (crRNA, purple line). The trans-activating CRISPR RNA (tracrRNA, blue line) is also required for this process (iii). b Representation of the chromosomally integrated reporter gfp-URA3 cassette to test the mutagenesis system. The genes gfp and URA3 are represented with green and yellow filled arrows, respectively. Thin arrows indicate the promoters tac (Ptac) and T7 (PT7), lollipops indicate terminators T0 and T1.
