Department of Biology and The Center for Genomics and Bioinformatics, Indiana University, Bloomington, IN 47405.
Proc Natl Acad Sci U S A. 2013 Nov 12;110(46):18584-9. doi: 10.1073/pnas.1309843110. Epub 2013 Oct 28.
Accurate transmission and expression of genetic information are crucial for the survival of all living organisms. Recently, the coupling of mutation accumulation experiments and next-generation sequencing has greatly expanded our knowledge of the genomic mutation rate in both prokaryotes and eukaryotes. However, because of their transient nature, transcription errors have proven extremely difficult to quantify, and current estimates of transcription fidelity are derived from artificial constructs applied to just a few organisms. Here we report a unique cDNA library preparation technique that allows error detection in natural transcripts at the transcriptome-wide level. Application of this method to the model organism Caenorhabditis elegans revealed a base misincorporation rate in mRNAs of ~4 × 10(-6) per site, with a very biased molecular spectrum. Because the proposed method is readily applicable to other organisms, this innovation provides unique opportunities for studying the incidence of transcription errors across the tree of life.
准确的遗传信息传递和表达对于所有生物的生存都是至关重要的。最近，突变积累实验和下一代测序的结合极大地扩展了我们对原核生物和真核生物基因组突变率的认识。然而，由于转录错误的瞬时性质，很难对其进行定量，目前对转录保真度的估计是从仅适用于少数几种生物体的人工构建体中得出的。在这里，我们报告了一种独特的 cDNA 文库制备技术，可在转录组水平上检测天然转录本中的错误。将该方法应用于模式生物秀丽隐杆线虫，揭示了 mRNA 中约每 4×10(-6)个碱基的碱基错配率，且分子谱非常偏向。由于所提出的方法易于应用于其他生物体，因此这项创新为研究生命之树中转录错误的发生率提供了独特的机会。
