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현정헌Jungheun Hyun

연세대학교

Exp. Mol. Med., 27 February 2024 | https://doi.org/10.1038/s12276-024-01164-8 Cost and time-efficient construction of a 3′-end mRNA library from unpurified bulk RNA in a single tube

Jungwon Choi ^1,4, Jungheun Hyun ^1,4, Jieun Hyun ^1,4, Jae-Hee Kim ¹, Ji Hyun Lee ^2,3,* and Duhee Bang ^1,*

¹Department of Chemistry, Yonsei University, Seoul, Republic of Korea.
²Department of Clinical Pharmacology and Therapeutics, College of Medicine, Kyung Hee University, Seoul, Republic of Korea.
³Department of Biomedical Science and Technology, Kyung Hee University, Seoul, Republic of Korea.
⁴These authors contributed equally: Jungwon Choi, Jungheun Hyun, Jieun Hyun

^*Corresponding authors: correspondence to Ji Hyun Lee or Duhee Bang

Abstract

The major drawbacks of RNA sequencing (RNA-seq), a remarkably accurate transcriptome profiling method, is its high cost and poor scalability. Here, we report a highly scalable and cost-effective method for transcriptomics profiling called Bulk transcriptOme profiling of cell Lysate in a single poT (BOLT-seq), which is performed using unpurified bulk 3′-end mRNA in crude cell lysates. During BOLT-seq, RNA/DNA hybrids are directly subjected to tagmentation, and second-strand cDNA synthesis and RNA purification are omitted, allowing libraries to be constructed in 2 h of hands-on time. BOLT-seq was successfully used to cluster small molecule drugs based on their mechanisms of action and intended targets. BOLT-seq competes effectively with alternative library construction and transcriptome profiling methods.

논문정보

형식Research article
게재일2024년 02월 (BRIC 등록일 2024-03-11)
연구진국내 연구진
분야 의약학 > 유전 및 유전체의학

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