Li Cheuk Yin, Liang Zhenghua, Hu Yaxin, Zhang Hongxia, Setiasabda Kharis Daniel, Li Jiawei, Ma Shaohua, Xia Xiaojun, Kuang Yi
Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China.
State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong 510060, China.
Mol Ther Nucleic Acids. 2022 Oct 12;30:300-310. doi: 10.1016/j.omtn.2022.10.003. eCollection 2022 Dec 13.
Synthetic mRNAs are rising rapidly as alternative therapeutic agents for delivery of proteins. However, the practical use of synthetic mRNAs has been restricted by their low cellular stability as well as poor protein production efficiency. The key roles of poly(A) tail on mRNA biology inspire us to explore the optimization of tail sequence to overcome the aforementioned limitations. Here, the systematic substitution of non-A nucleotides in the tails revealed that cytidine-containing tails can substantially enhance the protein production rate and duration of synthetic mRNAs both and . Such C-containing tails shield synthetic mRNAs from deadenylase CCR4-NOT transcription complex, as the catalytic CNOT proteins, especially CNOT6L and CNOT7, have lower efficiency in trimming of cytidine. Consistently, these enhancement effects of C-containing tails were observed on all synthetic mRNAs tested and were independent of transfection reagents and cell types. As the C-containing tails can be used along with other mRNA enhancement technologies to synergically boost protein production, we believe that these tails can be broadly used on synthetic mRNAs to directly promote their clinical applications.
作为蛋白质递送的替代治疗剂，合成mRNA正在迅速崛起。然而，合成mRNA的实际应用受到其低细胞稳定性以及低蛋白质生产效率的限制。聚（A）尾在mRNA生物学中的关键作用启发我们探索尾序列的优化，以克服上述限制。在这里，对尾巴中非A核苷酸的系统替换表明，含胞苷的尾巴可以显著提高合成mRNA在体外和体内的蛋白质产生速率和持续时间。这种含C的尾巴使合成mRNA免受去腺苷酸化酶CCR4-NOT转录复合物的影响，因为催化性CNOT蛋白，特别是CNOT6L和CNOT7，在修剪胞苷方面效率较低。一致地，在所有测试的合成mRNA上都观察到了这些含C尾巴的增强作用，并且与转染试剂和细胞类型无关。由于含C的尾巴可以与其他mRNA增强技术一起使用，以协同提高蛋白质产量，我们相信这些尾巴可以广泛用于合成mRNA，以直接促进其临床应用。
