Zhu Jun, Shendure Jay, Mitra Robi D, Church George M
Department of Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
Science. 2003 Aug 8;301(5634):836-8. doi: 10.1126/science.1085792.
Alternative pre-messenger RNA splicing is an important mechanism for generating protein diversity and may explain in part how mammalian complexity arises from a surprisingly small complement of genes. Here, we describe "digital polony exon profiling,"a single molecule-based technology for studying complex alternative pre-messenger RNA splicing. This technology allows researchers to monitor the combinatorial diversity of exon inclusion in individual transcripts. A minisequencing strategy provides single nucleotide resolution, and the digital nature of the technology allows quantitation of individual splicing variants. Digital polony exon profiling can be used to investigate the physiological and pathological roles of alternately spliced messenger RNAs, as well as the mechanisms by which these messenger RNAs are produced.
可变前体信使核糖核酸剪接是产生蛋白质多样性的重要机制，并且可能部分解释了哺乳动物的复杂性是如何从数量惊人少的基因补体中产生的。在此，我们描述了“数字克隆外显子分析”，这是一种基于单分子的技术，用于研究复杂的可变前体信使核糖核酸剪接。该技术使研究人员能够监测单个转录本中外显子包含的组合多样性。微测序策略提供单核苷酸分辨率，并且该技术的数字特性允许对单个剪接变体进行定量。数字克隆外显子分析可用于研究可变剪接信使核糖核酸的生理和病理作用，以及这些信使核糖核酸产生的机制。
