Translational modulation based on RNA-binding proteins can be used to construct artificial gene circuits, but RNA-binding proteins capable of regulating translation efficiently and orthogonally remain scarce. Here we report CARTRIDGE (Cas-Responsive Translational Regulation Integratable into Diverse Gene control) to repurpose Cas proteins as translational modulators in mammalian cells. We demonstrate that a set of Cas proteins efficiently and orthogonally repress or activate the translation of designed mRNAs that contain a Cas-binding RNA motif in the 5’-UTR. By linking multiple Cas-mediated translational modulators, we designed and built artificial circuits like logic gates, cascades, and half-subtractor circuits. Moreover, we show that various CRISPR-related technologies like anti-CRISPR and split-Cas9 platforms could be similarly repurposed to control translation. Coupling Cas-mediated translational and transcriptional regulation enhanced the complexity of synthetic circuits built by only introducing a few additional elements. Collectively, CARTRIDGE has enormous potential as a versatile molecular toolkit for mammalian synthetic biology.

基于 RNA 结合蛋白的翻译调节可用于构建人工基因回路，但能够有效和正交调节翻译的 RNA 结合蛋白仍然很少。在这里，我们报告了 CARTRIDGE（可整合到不同基因控制中的 Cas 响应性翻译调节）以将 Cas 蛋白重新用作哺乳动物细胞中的翻译调节剂。我们证明了一组 Cas 蛋白有效且正交地抑制或激活在 5'-UTR 中包含 Cas 结合 RNA 基序的设计 mRNA 的翻译。通过连接多个 Cas 介导的平移调制器，我们设计并构建了逻辑门、级联和半减法器电路等人工电路。而且，我们展示了各种 CRISPR 相关技术，如反 CRISPR 和分裂 Cas9 平台，可以类似地重新用于控制翻译。偶联 Cas 介导的翻译和转录调控提高了仅通过引入一些额外元素构建的合成电路的复杂性。总的来说，CARTRIDGE 作为哺乳动物合成生物学的多功能分子工具包具有巨大的潜力。