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Highly Multiplexed Subcellular RNA Sequencing in Situ

机译：高度复用的亚细胞RNA原位测序

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摘要

Understanding the spatial organization of gene expression with single-nucleotide resolution requires localizing the sequences of expressed RNA transcripts within a cell in situ. Here, we describe fluorescent in situ RNA sequencing (FISSEQ), in which stably cross-linked complementary DNA (cDNA) amplicons are sequenced within a biological sample. Using 30-base reads from 8102 genes in situ, we examined RNA expression and localization in human primary fibroblasts with a simulated wound-healing assay. FISSEQ is compatible with tissue sections and whole-mount embryos and reduces the limitations of optical resolution and noisy signals on single-molecule detection. Our platform enables massively parallel detection of genetic elements, including gene transcripts and molecular barcodes, and can be used to investigate cellular phenotype, gene regulation, and environment in situ.
机译：要了解具有单核苷酸分辨率的基因表达的空间组织，需要将表达的RNA转录物的序列原位定位在细胞内。在这里，我们描述了荧光原位RNA测序（FISSEQ），其中在生物样品中对稳定交联的互补DNA（cDNA）扩增子进行了测序。使用原位从8102个基因中读取的30个碱基，我们使用模拟的伤口愈合分析方法检测了人类原代成纤维细胞中的RNA表达和定位。 FISSEQ与组织切片和整装胚胎兼容，并减少了单分子检测中光学分辨率和噪声信号的限制。我们的平台能够大规模并行检测遗传元件，包括基因转录本和分子条形码，并可用于研究细胞表型，基因调控和原位环境。

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  • 来源
    《Science》 |2014年第6177期|1360-1363|共4页
  • 作者

    Je Hyuk Lee; Evan R. Daugharthy; Jonathan Scheiman; Reza Kalhor; Joyce L. Yang; Thomas C. Ferrante; Richard Terry; Sauveur S. F. Jeanty; Chao Li; Ryoji Amamoto; Derek T. Peters; Brian M. Turczyk; Adam H. Marblestone; Samuel A. Inverso; Amy Bernard; Prashant Mali; Xavier Rios; John Aach; George M. Church;

  • 作者单位

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA,Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA,Department of Genetics, Harvard Medical School, Boston, MA 02115, USA,Department of Systems Biology, Harvard Medical School Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA,Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA;

    Department of Stem Cell and Regenerative Biology, Harvard University, Boston, MA 02138, USA;

    Department of Stem Cell and Regenerative Biology, Harvard University, Boston, MA 02138, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA,Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA;

    Allen Institute for Brain Science, Seattle, WA 98103, USA;

    Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

    Wyss Institute, Harvard Medical School, Boston, MA 02115, USA,Department of Genetics, Harvard Medical School, Boston, MA 02115, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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  • 入库时间 2022-08-18 02:52:23

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