Artificial nondirectional site-specific recombination systems

Jun-Yi Wang; Yue-Yang Cao; Ya-Nan Chen; Xiao-Le Wu; Bo-Tao He; Si-Yu Zhu; Xiao Zhou; Yi Wu; Bing-Zhi Li; Ying-Jin Yuan

doi:10.1016/j.isci.2021.103716

Artificial nondirectional site-specific recombination systems

iScience. 2021 Dec 31;25(1):103716. doi: 10.1016/j.isci.2021.103716. eCollection 2022 Jan 21.

Authors

Jun-Yi Wang^{1

2}, Yue-Yang Cao^{1

2}, Ya-Nan Chen^{1

2}, Xiao-Le Wu^{1

2}, Bo-Tao He^{1

2}, Si-Yu Zhu^{1

2}, Xiao Zhou^{1

2}, Yi Wu^{1

2}, Bing-Zhi Li^{1

2}, Ying-Jin Yuan^{1

2}

Affiliations

¹ Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
² Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin 300072, China.

Abstract

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/voxsym1-4 and Dre/roxsym1-2. 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 simultaneously 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.

Keywords: Bioengineering; Biological sciences; Biological sciences research methodologies; Biotechnology; Genetic engineering; Synthetic biology.