Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 117, Issue 6, Pages 2886-2893Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1919800117
Keywords
single cell; RNA-seq; Tn5 transposase
Categories
Funding
- National Natural Science Foundation of China [21675098, 21525521]
- Ministry of Science and Technology of China [2018YFA0800200, 2018YFA0108100, 2018YFC1002300]
- Beijing Brain Initiative [Z181100001518004]
- Beijing Advanced Innovation Center for Structural Biology
- Beijing Advanced Innovation Center for Genomics
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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.
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