Designing logical codon reassignment - Expanding the chemistry in biology.
Designing logical codon reassignment - Expanding the chemistry in biology.
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DOI：
10.1039/c4sc01534g
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发表时间：
2015-01-01
期刊：
影响因子：
8.4
通讯作者：
Davis BG
中科院分区：
文献类型：
--
作者：
Dumas A;Lercher L;Spicer CD;Davis BG
这篇综述使通过规范密码子将尿酸结合到蛋白质中的不同系统设计合理化。在过去的十年里，对非天然氨基酸(UAA)进行基因编码的能力得到了迅速发展。UAAs的程序性结合依赖于用氨基-酰基tRNA合成酶/tRNA(AARS/tRNA)对重新分配或抑制规范密码子，选择性地选择UAA。为了实现选择性掺入，AAR应该对设计的tRNA和UAA具有选择性，而不是内源氨基酸和tRNAs。通过将AARS/tRNA对从另一个王国转移到目标生物体，以及随后的AARS进化来获得对所需UAA的增强选择性，实现了增强的选择性。今天，已有超过150种非规范氨基酸使用这种方法被结合。这使得在从原核生物、酵母和哺乳动物细胞系到整个动物的有机体中，蛋白质中引入了各种各样的结构，使蛋白质功能的研究达到了以前无法实现的水平。虽然到目前为止，大多数研究都集中在抑制“无义”密码子上，但最近的发展开始打开四重密码子解码的可能性，并更有选择性地重新分配有义密码子，为整合多种氨基酸提供了一个潜在的强大工具。在此，我们旨在对UAA掺入方法进行重点综述，特别是对已开发或开发的不同tRNA合成酶/tRNA对，重点放在已被掺入的不同UAA结构以及此类系统设计和未来创建背后的逻辑。我们希望，这将有助于合理化系统的设计，纳入未探索的非天然氨基酸，以及那些已知的新应用。
This review rationalizes the varied designs of systems for incorporation of UAAs into proteins via canonical codons. Over the last decade, the ability to genetically encode unnatural amino acids (UAAs) has evolved rapidly. The programmed incorporation of UAAs into recombinant proteins relies on the reassignment or suppression of canonical codons with an amino-acyl tRNA synthetase/tRNA (aaRS/tRNA) pair, selective for the UAA of choice. In order to achieve selective incorporation, the aaRS should be selective for the designed tRNA and UAA over the endogenous amino acids and tRNAs. Enhanced selectivity has been achieved by transferring an aaRS/tRNA pair from another kingdom to the organism of interest, and subsequent aaRS evolution to acquire enhanced selectivity for the desired UAA. Today, over 150 non-canonical amino acids have been incorporated using such methods. This enables the introduction of a large variety of structures into proteins, in organisms ranging from prokaryote, yeast and mammalian cells lines to whole animals, enabling the study of protein function at a level that could not previously be achieved. While most research to date has focused on the suppression of ‘non-sense’ codons, recent developments are beginning to open up the possibility of quadruplet codon decoding and the more selective reassignment of sense codons, offering a potentially powerful tool for incorporating multiple amino acids. Here, we aim to provide a focused review of methods for UAA incorporation with an emphasis in particular on the different tRNA synthetase/tRNA pairs exploited or developed, focusing upon the different UAA structures that have been incorporated and the logic behind the design and future creation of such systems. Our hope is that this will help rationalize the design of systems for incorporation of unexplored unnatural amino acids, as well as novel applications for those already known.