Reverse transcriptases (RTs) are enzymes that can generate a complementary strand of DNA (cDNA) from RNA. Coupled with PCR, RTs have been widely used to detect RNAs and to clone expressed genes. Classical retroviral RTs have been improved by protein engineering. These enzymes and newly characterized RTs are key elements in the development of next-generation sequencing techniques that are now being applied to the study of transcriptomics. In addition, engineered RTs fused to a CRISPR/Cas9 nickase have recently shown great potential as tools to manipulate eukaryotic genomes. In this review, we discuss the properties and uses of wild type and engineered RTs in biotechnological applications, from conventional RT-PCR to recently introduced prime editing.

逆转录酶 (RT) 是可以从 RNA 生成互补 DNA (cDNA) 链的酶。与 PCR 相结合，RT 已被广泛用于检测 RNA 和克隆表达的基因。经典的逆转录病毒 RT 已通过蛋白质工程得到改进。这些酶和新表征的 RT 是开发下一代测序技术的关键要素，这些技术现在正应用于转录组学研究。此外，与 CRISPR/Cas9 切口酶融合的工程化 RT 最近显示出作为操纵真核基因组工具的巨大潜力。在这篇综述中，我们讨论了野生型和工程化 RT 在生物技术应用中的特性和用途，从传统的 RT-PCR 到最近引入的主要编辑。