Artificial nondirectional site-specific recombination systems

Site-specific recombination systems (SRSs) are widely used in studies on synthetic biology and related disciplines. Nondirectional SRSs can randomly trigger excision, integration, reversal, and translocation, which are effective tools to achieve large-scale genome recombination. In this study, we designed 6 new nondirectional SRSs named Vika/voxsyml-4 and Dre/roxsyml-2. All 6 artificial nondirectional SRSs were able to generate random deletion and inversion in Saccharomyces cerevisiae. Moreover, all six SRSs were orthogonal to Cre/loxPsym. The pairwise orthogonal nondirected SRSs can sime_aneously initiate large-scale and independent gene recombination in two different regions of the genome, which could not be accomplished using previous orthogonal systems. These SRSs were found to be robust while working in the cells at different growth stages, as well as in the different spatial structure of the chromosome. These artificial pairwise orthogonal nondirected SRSs offer newfound potential for site-specific recombination in synthetic biology.

Automated summary

Site-specific recombination systems (SRSs) play an important role in synthetic biology research. Nondirectional SRSs can randomly trigger genome recombination, and in this study, 6 new nondirectional SRSs were designed and successfully generated random recombination in Saccharomyces cerevisiae. These SRSs can independently initiate large-scale gene recombination in different regions of the genome, and they exhibit stability in different cell growth stages and chromosome structures.