Engineering aminoacyl-tRNA synthetases for use in synthetic biology.

Engineering aminoacyl-tRNA synthetases for use in synthetic biology.
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DOI:
10.1016/bs.enz.2020.06.004
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发表时间:
2020
期刊:
The Enzymes
影响因子:
--
通讯作者:
Söll D
Söll D
中科院分区:
其他
文献类型:
--
作者:
Krahn N;Tharp JM;Crnković A;Söll D

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在合成生物学的广泛领域内,遗传密码扩展(GCE)技术能够创建具有扩展的氨基酸组的蛋白质。这对于治疗学、生物修复和生物催化的应用可能是无价的。GCE的核心是氨酰-tRNA合成酶(aaRS),因为它们将非规范氨基酸(ncAA)连接到其同源tRNA上,允许ncAA掺入核糖体上的蛋白质中。ncAA-酰化aaRS及其tRNA不应与宿主中的20种天然aaRS和tRNA交叉反应,即,它们需要充当正交翻译系统。目前所有的正交aaRS·tRNA对都是从天然存在的分子中改造而来的,以改变aaRS的氨基酸特异性或将tRNA分配给所选择的释放密码子。在这里,我们讨论了正交性在GCE中的重要性,用于创建设计师aaRS和tRNA的实验室技术,并提供了用于GCE目的的正交aaRS·tRNA对的概述。
Within the broad field of synthetic biology, genetic code expansion (GCE) techniques enable creation of proteins with an expanded set of amino acids. This may be invaluable for applications in therapeutics, bioremediation, and biocatalysis. Central to GCE are aminoacyl-tRNA synthetases (aaRSs) as they link a non-canonical amino acid (ncAA) to their cognate tRNA, allowing ncAA incorporation into proteins on the ribosome. The ncAA-acylating aaRSs and their tRNAs should not cross-react with 20 natural aaRSs and tRNAs in the host, i.e., they need to function as an orthogonal translating system. All current orthogonal aaRS•tRNA pairs have been engineered from naturally occurring molecules to change the aaRS’s amino acid specificity or assign the tRNA to a liberated codon of choice. Here we discuss the importance of orthogonality in GCE, laboratory techniques employed to create designer aaRSs and tRNAs, and provide an overview of orthogonal aaRS•tRNA pairs for GCE purposes.
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