Split-TALE: A TALE-Based Two-Component System for Synthetic Biology Applications in Planta

被引:15
作者
Schreiber, Tom [1 ]
Prange, Anja [1 ]
Hoppe, Tina [2 ]
Tissier, Alain [1 ]
机构
[1] Leibniz Inst Plant Biochem, Dept Cell & Metab Biol, D-06120 Halle, Saale, Germany
[2] Martin Luther Univ Halle Wittenberg, Inst Biol, Dept Genet, D-06120 Halle, Saale, Germany
关键词
CRYSTAL-STRUCTURE; TRANSCRIPTION FACTOR; PROTEIN; DNA; EXPRESSION; EFFECTORS; GENE; ACTIVATOR; SPECIFICITY; RECOGNITION;
D O I
10.1104/pp.18.01218
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Transcription activator-like effectors (TALEs) are bacterial Type-III effector proteins from phytopathogenic Xanthomonas species that act as transcription factors in plants. The modular DNA-binding domain of TALEs can be reprogrammed to target nearly any DNA sequence. Here, we designed and optimized a two-component AND-gate system for synthetic circuits in plants based on TALEs. In this system, named split-TALE (sTALE), the TALE DNA binding domain and the transcription activation domain are separated and each fused to protein interacting domains. Physical interaction of interacting domains leads to TALE-reconstitution and can be monitored by reporter gene induction. This setup was used for optimization of the sTALE scaffolds, which result in an AND-gate system with an improved signal-to-noise ratio. We also provide a toolkit of ready-to-use vectors and single modules compatible with Golden Gate cloning and MoClo syntax. In addition to its implementation in synthetic regulatory circuits, the sTALE system allows the analysis of protein-protein interactions in planta.
引用
收藏
页码:1001 / 1012
页数:12
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