Engineered ribosomes with tethered subunits for expanding biological function
Engineered ribosomes with tethered subunits for expanding biological function
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DOI：
10.1038/s41467-019-11427-y
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发表时间：
2019-09-02
影响因子：
16.6
通讯作者：
Jewett, Michael C.
中科院分区：
文献类型：
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作者：
Carlson, Erik D.;D'Aquino, Anne E.;Jewett, Michael C.
Ribo-T是一种核糖体，具有共价束缚的亚基，其中核心16S和23S核糖体RNA形成单个嵌合分子。 Ribo-T使细胞中功能性正交核糖体mRNA系统成为可能。不幸的是，由于其原始版本的活性较低，因此使用Ribo-T受到限制。在这里，为了克服这一限制，我们使用一种进化方法来选择能够支持更快的细胞生长和蛋白质表达增加的新链球设计。此外，我们进化了与自然核糖体和mRNA并行作用的新的正交核糖-T/mRNA对，从而增加了正交蛋白表达的效率。具有优化设计的RIBO-T能够合成各种蛋白质，还可以将多种非典型氨基酸掺入合成的多肽中。增强的Ribo-T设计对于探索核糖体和工程核糖体的功能较低的功能应具有改变的催化特性，这应该很有用。
Ribo-T is a ribosome with covalently tethered subunits where core 16S and 23S ribosomal RNAs form a single chimeric molecule. Ribo-T makes possible a functionally orthogonal ribosome-mRNA system in cells. Unfortunately, use of Ribo-T has been limited because of low activity of its original version. Here, to overcome this limitation, we use an evolutionary approach to select new tether designs that are capable of supporting faster cell growth and increased protein expression. Further, we evolve new orthogonal Ribo-T/mRNA pairs that function in parallel with, but independent of, natural ribosomes and mRNAs, increasing the efficiency of orthogonal protein expression. The Ribo-T with optimized designs is able to synthesize a diverse set of proteins, and can also incorporate multiple non-canonical amino acids into synthesized polypeptides. The enhanced Ribo-T designs should be useful for exploring poorly understood functions of the ribosome and engineering ribosomes with altered catalytic properties.