Highly Multiplexed Subcellular RNA Sequencing in Situ

被引：752

作者：

Lee, Je Hyuk ^{[1
,2
]}

Daugharthy, Evan R. ^{[1
,2
,4
]}

Scheiman, Jonathan ^{[1
,2
]}

Kalhor, Reza ^{[2
]}

Yang, Joyce L. ^{[2
]}

Ferrante, Thomas C. ^{[1
]}

Terry, Richard ^{[1
]}

Jeanty, Sauveur S. F. ^{[1
]}

Li, Chao ^{[1
]}

Amamoto, Ryoji ^{[3
]}

Peters, Derek T. ^{[3
]}

Turczyk, Brian M. ^{[1
]}

Marblestone, Adam H. ^{[1
,2
]}

Inverso, Samuel A. ^{[1
]}

Bernard, Amy ^{[5
]}

Mali, Prashant ^{[2
]}

Rios, Xavier ^{[2
]}

Aach, John ^{[2
]}

Church, George M. ^{[1
,2
]}

机构：

[1] Harvard Univ, Sch Med, Wyss Inst, Boston, MA 02115 USA

[2] Harvard Univ, Dept Genet, Sch Med, Boston, MA 02115 USA

[3] Harvard Univ, Dept Stem Cell & Regenerat Biol, Boston, MA 02138 USA

[4] Harvard Univ, Dept Syst Biol, Sch Med, Boston, MA 02115 USA

[5] Allen Inst Brain Sci, Seattle, WA 98103 USA

来源：

SCIENCE | 2014年 / 343卷 / 6177期

关键词：

GENE-EXPRESSION; TRANSCRIPTS; REVEALS; TISSUE;

D O I：

10.1126/science.1250212

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

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.

引用

页码：1360 / 1363

页数：4

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