Kwak Yeonui, Kwak Hojoong
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA.
Graduate Field of Genetics, Genomics, and Developmental Biology, Cornell University, Ithaca, NY, USA.
Methods Mol Biol. 2022;2404:281-298. doi: 10.1007/978-1-0716-1851-6_15.
Polyadenylation and deadenylation of mRNA are major RNA modifications associated with nucleus-to-cytoplasm translocation, mRNA stability, translation efficiency, and mRNA decay pathways. Our current knowledge of polyadenylation and deadenylation has been expanded due to recent advances in transcriptome-wide poly(A) tail length assays. Whereas these methods measure poly(A) length by quantifying the adenine (A) base stretch at the 3' end of mRNA, we developed a more cost-efficient technique that does not rely on A-base counting, called tail-end-displacement sequencing (TED-seq). Through sequencing highly size-selected 3' RNA fragments including the poly(A) tail pieces, TED-seq provides accurate measure of transcriptome-wide poly(A)-tail lengths in high resolution, economically suitable for larger scale analysis under various biologically transitional contexts.
mRNA的聚腺苷酸化和去腺苷酸化是与细胞核到细胞质转运、mRNA稳定性、翻译效率以及mRNA降解途径相关的主要RNA修饰。由于转录组范围的聚(A)尾长度测定技术的最新进展,我们目前对聚腺苷酸化和去腺苷酸化的认识得到了扩展。虽然这些方法通过量化mRNA 3'端的腺嘌呤(A)碱基延伸来测量聚(A)长度,但我们开发了一种更具成本效益的技术,该技术不依赖于A碱基计数,称为尾端位移测序(TED-seq)。通过对包括聚(A)尾片段在内的高度大小选择的3'RNA片段进行测序,TED-seq以高分辨率提供转录组范围聚(A)尾长度的准确测量,在经济上适合于各种生物学过渡背景下的大规模分析。