德国Max Delbrück分子医学研究所Nikolaus Rajewsky研究团队取得一项新进展，他们研发出一个名为FLAM-seq（全长poly(A)和mRNA测序）的方法，可对全长mRNA进行测序并据此揭示出poly(A)尾长度调控原则。该研究于2019年9月发表于国际一流学术期刊《自然—方法学》上。

FLAM-seq是一种快速简便的方法，可对整个mRNA进行高质量测序。研究人员报道了一种互补的DNA文库制备方法，该方法与单分子测序相结合，可以进行FLAM-seq。使用人类细胞系、大脑类器官和秀丽隐杆线虫，研究人员发现FLAM-seq为每个样品提供数千种不同基因的高质量全长mRNA序列。研究人员也发现3'端非翻译区长度与poly(A)尾长相关，即相同基因的可变多腺苷酸化位点和可变启动子与不同的尾长相关，并且尾部含有大量胞嘧啶。

据介绍，虽然信使RNA是理解生命的关键分子，但直到现在还没有方法来确定内源mRNA的全长序列，包括它们的poly(A)尾。此外，虽然非A核苷酸可以掺入poly(A)尾中，但也没有方法可以精确地对它们进行测序。

附：英文原文

Title: FLAM-seq: full-length mRNA sequencing reveals principles of poly(A) tail length control

Author: Ivano Legnini, Jonathan Alles, Nikos Karaiskos, Salah Ayoub, Nikolaus Rajewsky

Issue&Volume: Volume 16 Issue 9

Abstract: Although messenger RNAs are key molecules for understanding life, until now, no method has existed to determine the full-length sequence of endogenous mRNAs including their poly(A) tails. Moreover, although non-A nucleotides can be incorporated in poly(A) tails, there also exists no method to accurately sequence them. Here, we present full-length poly(A) and mRNA sequencing (FLAM-seq), a rapid and simple method for high-quality sequencing of entire mRNAs. We report a complementary DNA library preparation method coupled to single-molecule sequencing to perform FLAM-seq. Using human cell lines, brain organoids and Caenorhabditis elegans we show that FLAM-seq delivers high-quality full-length mRNA sequences for thousands of different genes per sample. We find that 3′ untranslated region length is correlated with poly(A) tail length, that alternative polyadenylation sites and alternative promoters for the same gene are linked to different tail lengths, and that tails contain a substantial number of cytosines.

DOI: 10.1038/s41592-019-0503-y

Source: https://www.nature.com/articles/s41592-019-0503-y