Pietschmann Max, Tempel Gregor, Halladjian Maral, Krogh Nicolai, Nielsen Henrik
Department of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2021;2167:271-285. doi: 10.1007/978-1-0716-0716-9_15.
A lariat cap is a naturally occurring substitute of a conventional mRNA cap and is found in a particular genomic setting in a few eukaryotic microorganisms. It is installed by the lariat capping ribozyme acting in cis. In principle, any RNA molecule in any organism can be equipped with a lariat cap in vivo when expressed downstream of a lariat capping ribozyme. Lariat capping is thus a versatile tool for studying the importance of the 5' end structure of RNA molecules. In this chapter, we present protocols to validate the presence of the lariat cap and measure the efficiency of in vivo cleavage by the lariat capping ribozyme.
套索帽是传统mRNA帽的天然替代品，在一些真核微生物的特定基因组环境中被发现。它由顺式作用的套索帽核酶安装。原则上，任何生物体中的任何RNA分子在套索帽核酶下游表达时，都可以在体内配备套索帽。因此，套索帽加帽是研究RNA分子5'端结构重要性的一种通用工具。在本章中，我们介绍了验证套索帽存在并测量套索帽核酶体内切割效率的实验方案。
