RNA sequencing by direct tagmentation of RNA/DNA hybrids

被引：86

作者：

Di, Lin ^{[1
]}

Fu, Yusi ^{[1
,12
]}

Sun, Yue ^{[1
]}

Li, Jie ^{[2
,3
]}

Liu, Lu ^{[2
,3
]}

Yao, Jiacheng ^{[2
,3
]}

Wang, Guanbo ^{[4
,5
]}

Wu, Yalei ^{[6
]}

Lao, Kaiqin ^{[6
]}

Lee, Raymond W. ^{[6
]}

Zheng, Genhua ^{[6
]}

Xu, Jun ^{[7
]}

Oh, Juntaek ^{[7
]}

Wang, Dong ^{[7
]}

Xie, X. Sunney ^{[1
]}

Huang, Yanyi ^{[1
,5
,8
,9
,10
]}

Wang, Jianbin ^{[2
,3
,10
,11
]}

机构：

[1] Peking Univ, Beijing Adv Innovat Ctr Genom ICG, Sch Life Sci, Biomed Pioneering Innovat Ctr BIOPIC, Beijing 100871, Peoples R China

[2] Tsinghua Univ, Sch Life Sci, Beijing 100084, Peoples R China

[3] Tsinghua Univ, Tsinghua Peking Ctr Life Sci, Beijing 100084, Peoples R China

[4] Nanjing Normal Univ, Sch Chem & Mat Sci, Nanjing 210046, Peoples R China

[5] Shenzhen Bay Lab, Inst Cell Anal, Shenzhen 518132, Peoples R China

[6] XGen US Co, San Francisco, CA 94080 USA

[7] Univ Calif San Diego, Dept Cellular & Mol Med, Skaggs Sch Pharm & Pharmaceut Sci, La Jolla, CA 92093 USA

[8] Peking Univ, Coll Engn, Beijing 100871, Peoples R China

[9] Peking Univ, Coll Chem & Mol Engn, Beijing 100871, Peoples R China

[10] CIBR, Beijing 102206, Peoples R China

[11] Tsinghua Univ, Beijing Adv Innovat Ctr Struct Biol, Beijing 100084, Peoples R China

[12] Baylor Coll Med, Dept Mol & Human Genet, Houston, TX 77030 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2020年 / 117卷 / 06期

基金：

中国国家自然科学基金;

关键词：

single cell; RNA-seq; Tn5; transposase; TN5; TRANSPOSASE; SINGLE CELLS; SEQ; AMPLIFICATION; DNA;

D O I：

10.1073/pnas.1919800117

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：

07 ; 0710 ; 09 ;

摘要：

Transcriptome profiling by RNA sequencing (RNA-seq) has been widely used to characterize cellular status, but it relies on second-strand complementary DNA (cDNA) synthesis to generate initial material for library preparation. Here we use bacterial transposase Tn5, which has been increasingly used in various high-throughput DNA analyses, to construct RNA-seq libraries without second-strand synthesis. We show that Tn5 transposome can randomly bind RNA/DNA heteroduplexes and add sequencing adapters onto RNA directly after reverse transcription. This method, Sequencing HEteRo RNA-DNA-hYbrid (SHERRY), is versatile and scalable. SHERRY accepts a wide range of starting materials, from bulk RNA to single cells. SHERRY offers a greatly simplified protocol and produces results with higher reproducibility and GC uniformity compared with prevailing RNA-seq methods.

引用

页码：2886 / 2893

页数：8

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