PAIso-seq

  • 文章类型: Journal Article
    Poly(A)尾被添加到大多数真核mRNA中,并具有必要的调节功能。然而,由于其均聚性质,poly(A)尾部的序列信息在转录组测量研究中很难获得。在这一章中,我们描述了poly(A)包容性全长RNA同工型测序(PAIso-seq)的详细程序,一种基于PacBioHiFi测序平台的方法,可以从低至纳克水平的总RNA中测量转录组范围的poly(A)尾部。可以与全长cDNA一起获得聚(A)尾的精确长度和碱基组成。
    Poly(A) tails are added to most eukaryotic mRNA and have essential regulatory functions. However, due to its homopolymeric nature, the sequence information in poly(A) tails is challenging to obtain in transcriptome measurement studies. In this chapter, we describe the detailed procedures of poly(A) inclusive full-length RNA isoform-sequencing (PAIso-seq), a method that can measure transcriptome-wide poly(A) tails from as low as nanogram level of total RNA based on the PacBio HiFi sequencing platform. The accurate length and base composition of poly(A) tails can be obtained along with the full-length cDNA.
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  • 文章类型: Review
    During mammalian oocyte-to-embryo transition, before zygotic genome activation, the transcription in oocytes and embryos is silenced, so the post-transcriptional regulation of mRNA plays an essential role in this process. Poly(A) tail is an important post-transcriptional modification that affects mRNA metabolism and translation efficiency. With the development of sequencing technology and analysis tools, especially the methods based on third-generation sequencing technology, the length and composition of poly(A) tails can be accurately measured, greatly expanding our understanding of poly(A) tails in mammalian early embryonic development. This review focuses on the achievements of poly(A) tail sequencing methods and the research progress of poly(A) tail in regulating oocyte-to-embryo transition, discussing the future applications for the investigation of mammalian early embryonic development and infertility related diseases.
    在哺乳动物卵子向胚胎转变过程中,卵子和胚胎中的转录在合子基因组激活之前都是沉默的,因此mRNA转录后修饰在此过程发挥着重要的作用。poly(A)尾巴是影响mRNA命运和翻译效率的一种重要的转录后修饰。随着测序技术和分析工具的进步,尤其是三代测序技术的发展,poly(A)尾的长度和组成能够被精确测量,极大地拓展了人们对于poly(A)尾在哺乳动物早期胚胎发育过程中的认识。本文对poly(A)尾研究方法的发展及其在卵子向胚胎转变中的研究进展进行评述,以期为哺乳动物早期胚胎发育和不孕不育相关疾病的研究带来新的思路。.
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  • 文章类型: Journal Article
    理解生命开始时的一个非凡事件,卵母细胞到胚胎的过渡(OET),仍然难以捉摸,尤其是在人类中。使用新开发的技术,Liuetal.表明人类母体mRNA在OET过程中经历了全局poly(A)尾介导的重塑,确定了所涉及的酶,并证明了重塑胚胎卵裂的必要性。
    Understanding a remarkable event at the start of life, the oocyte-to-embryo transition (OET), has remained elusive, especially in humans. Using newly developed techniques, Liu et al. showed that human maternal mRNAs undergo global poly(A) tail-mediated remodeling during OET, identified the enzymes involved, and demonstrated the essentiality of remodeling for embryo cleavage.
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