School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, China.
Methods Mol Biol. 2024;2760:95-114. doi: 10.1007/978-1-0716-3658-9_6.
We describe a new way to trigger mRNA degradation in Saccharomyces cerevisiae synthetic gene circuits. Our method demands to modify either the 5'- or the 3'-UTR that flanks a target gene with elements from the pre-crRNA of type V Cas12a proteins and expresses a DNase-deficient Cas12a (dCas12a). dCas12a recognizes and cleaves the pre-crRNA motifs on mRNA sequences. Our tool does not require complex engineering operations and permits an efficient control of protein expression via mRNA degradation.
我们描述了一种在酿酒酵母合成基因回路中触发 mRNA 降解的新方法。我们的方法要求通过来自 V 型 Cas12a 蛋白的 pre-crRNA 元件修饰侧翼靶基因的 5' 或 3'UTR，并表达无 DNA 酶活性的 Cas12a（dCas12a）。dCas12a 识别并切割 mRNA 序列上的 pre-crRNA 基序。我们的工具不需要复杂的工程操作，并允许通过 mRNA 降解来有效控制蛋白质表达。
