Figure 1.

Method for expression of multiple genes. (a) Overview of the CRISPR/Cas9‐RNA interference workflow for expressing multiple genes. First, expression constructs are assembled using USER cloning‐ligation‐PCR. The promoter and terminator are chosen to obtain the desired gene expression level. In the next step, the expression constructs are transformed into Cas9‐expressing yeast strain, along with upstream and downstream repair fragments and a selective marker. (b) Fluorescent cytometry analysis of four Saccharomyces cerevisiae strains, where genes encoding for red (RFP), cyan (CFP), and yellow (YFP) fluorescent proteins were expressed under control of promoters with different strengths [Colour figure can be viewed at wileyonlinelibrary.com]