Nucleic Acid Chemistry and Engineering Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904 0495, Japan.
Methods. 2019 May 15;161:41-45. doi: 10.1016/j.ymeth.2019.02.001. Epub 2019 Feb 6.
A large number of catalytic RNAs, or ribozymes, have been identified in the genomes of various organisms and viruses. Ribozymes are involved in biological processes such as regulation of gene expression and viral replication, but biological roles of many ribozymes still remain unknown. Ribozymes have also inspired researchers to engineer synthetic ribozymes that function as sensors or gene switches. To gain deeper understanding of the sequence-function relationship of ribozymes and to efficiently engineer synthetic ribozymes, a large number of ribozyme variants need to be examined which was limited to hundreds of sequences by Sanger sequencing. The advent of high-throughput sequencing technologies, however, has allowed us to sequence millions of ribozyme sequences at low cost. This review focuses on the recent applications of high-throughput sequencing to both characterize and engineer ribozymes, to highlight how the large-scale sequence data can advance ribozyme research and engineering.
大量的催化 RNA（或核酶）已在各种生物体和病毒的基因组中被鉴定出来。核酶参与基因表达调控和病毒复制等生物过程，但许多核酶的生物学功能仍然未知。核酶也启发了研究人员设计具有传感器或基因开关功能的合成核酶。为了更深入地了解核酶的序列-功能关系，并有效地设计合成核酶，需要对大量的核酶变体进行研究，而桑格测序技术将这个数量限制在数百个序列以内。然而，高通量测序技术的出现使得我们能够以低成本对数百万个核酶序列进行测序。本综述重点介绍了高通量测序在核酶的表征和工程设计方面的最新应用，强调了大规模序列数据如何推动核酶的研究和工程设计。
