Institut de biologie moléculaire des plantes, CNRS, Université de Strasbourg, Strasbourg, France.
Methods Mol Biol. 2024;2723:233-252. doi: 10.1007/978-1-0716-3481-3_14.
Deadenylation is a major process that regulates gene expression by shaping the length of mRNA poly(A) tails. Deadenylation is controlled by factors in trans that recruit or impede deadenylases, by the incorporation of non-adenosines during poly(A) tail synthesis, and by the posttranscriptional addition of 3' nucleotides to poly(A) tails. Deciphering the regulation of poly(A) tail shortening requires both transcriptome-wide approaches and more targeted methodologies, allowing deep analyses of specific mRNAs. In this chapter, we present Nano3'RACE, a nanopore-based cDNA sequencing method that allows in-depth analysis to precisely measure poly(A) tail length and detect 3' terminal nucleotide addition, such as uridylation, for mRNAs of interest.
脱腺苷酸化是一种主要的调控基因表达的过程，通过改变 mRNA 多聚（A）尾的长度来实现。脱腺苷酸化受顺式作用因子的调控，这些因子招募或阻碍脱腺苷酸酶，通过在多聚（A）尾合成过程中掺入非腺苷酸，以及通过转录后在多聚（A）尾上添加 3' 核苷酸来实现。解析多聚（A）尾缩短的调控需要转录组范围的方法和更有针对性的方法，以便对特定的 mRNAs 进行深入分析。在本章中，我们介绍了 Nano3'RACE，这是一种基于纳米孔的 cDNA 测序方法，允许进行深入分析，以精确测量多聚（A）尾的长度，并检测感兴趣的 mRNAs 的 3' 末端核苷酸添加，如尿苷酸化。
