To spatially control biochemical functions at specific sites within a genome, we have engineered a synthetic switch that activates when bound to its DNA target site. The system uses two CRISPR–Cas complexes to colocalize components of a de novo-designed protein switch (Co-LOCKR) to adjacent sites in the genome. Colocalization triggers a conformational change in the switch from an inactive closed state to an active open state with an exposed functional peptide. We prototype the system in yeast and demonstrate that DNA binding triggers activation of the switch, recruitment of a transcription factor, and expression of a downstream reporter gene. This DNA-triggered Co-LOCKR switch provides a platform to engineer sophisticated functions that should only be executed at a specific target site within the genome, with potential applications in a wide range of synthetic systems including epigenetic regulation, imaging, and genetic logic circuits.

中文翻译：

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使用共定位依赖的蛋白质开关有条件地招募 DNA 结合的 CRISPR-Cas 复合物。


为了在空间上控制基因组内特定位点的生化功能，我们设计了一种合成开关，当结合到其 DNA 靶位点时，该开关就会激活。该系统使用两个 CRISPR-Cas 复合物将从头设计的蛋白质开关 (Co-LOCKR) 的组件共定位到基因组中的相邻位点。共定位触发了与暴露的功能肽从非活性闭合状态到活性开放状态的构象变化。我们在酵母中建立了该系统原型，并证明 DNA 结合会触发开关的激活、转录因子的募集以及下游报告基因的表达。这种 DNA 触发的 Co-LOCKR 开关提供了一个平台来设计复杂的功能，这些功能只能在基因组内的特定目标位点执行，在广泛的合成系统中具有潜在的应用，包括表观遗传调控、成像和遗传逻辑电路。