강태준 (한국생명공학연구원) | 한빛사논문 > 한빛사

한빛사논문

조회551

한국생명공학연구원

CV File 1 MB

Nat. Biotechnol., 07 October 2024 | https://doi.org/10.1038/s41587-024-02426-6 Amplifying mutational profiling of extracellular vesicle mRNA with SCOPE

Jayeon Song ^1,2,3,4, Mi Hyeon Cho ^1,2, Hayoung Cho ¹, Younseong Song ⁵, Sung Woon Lee ⁶, Ho Chul Nam⁶, Tae Ho Yoon ⁶, Jong Cheol Shin ⁶, Jae-Sang Hong ^1,2, Yejin Kim ⁷, Emil Ekanayake ⁸, Jueun Jeon ^1,2, Dong Gil You ^1,8, Sung Gap Im ⁵, Gyu-Seog Choi⁹, Jun Seok Park ⁹, Bob C. Carter ⁸, Leonora Balaj ⁸, An Na Seo ¹⁰, Miles A. Miller ^1,2, Soo Yeun Park ⁹, Taejoon Kang ^4,11,*, Cesar M. Castro ^1,12,* & Hakho Lee ^1,2,13,*

¹Center for Systems Biology, Massachusetts General Hospital Research Institute, Boston, MA, USA.
²Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
³Department of Forensic Sciences, Sungkyunkwan University, Suwon, Republic of Korea.
⁴School of Pharmacy, Sungkyunkwan University, Suwon, Republic of Korea.
⁵Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.
⁶RevoSketch, Inc., Daejeon, Republic of Korea.
⁷Research Institute of Aging and Metabolism, Kyungpook National University, Daegu, Republic of Korea.
⁸Department of Neurosurgery, Massachusetts General Hospital, Boston, MA, USA.
⁹Colorectal Cancer Center, Kyungpook National University Chilgok Hospital, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
¹⁰Department of Pathology, School of Medicine, Kyungpook National University, Kyungpook National University Chilgok Hospital, Daegu, Republic of Korea.
¹¹Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea.
¹²Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
¹³Center for Nanomedicine, Institute for Basic Science (IBS), Seoul, Republic of Korea.

^*Corresponding authors: correspondence to Taejoon Kang, Cesar M. Castro or Hakho Lee

Abstract

Sequencing of messenger RNA (mRNA) found in extracellular vesicles (EVs) in liquid biopsies can provide clinical information such as somatic mutations, resistance profiles and tumor recurrence. Despite this, EV mRNA remains underused due to its low abundance in liquid biopsies, and large sample volumes or specialized techniques for analysis are required. Here we introduce Self-amplified and CRISPR-aided Operation to Profile EVs (SCOPE), a platform for EV mRNA detection. SCOPE leverages CRISPR-mediated recognition of target RNA using Cas13 to initiate replication and signal amplification, achieving a sub-attomolar detection limit while maintaining single-nucleotide resolution. As a proof of concept, we designed probes for key mutations in KRAS, BRAF, EGFR and IDH1 genes, optimized protocols for single-pot assays and implemented an automated device for multi-sample detection. We validated SCOPE’s ability to detect early-stage lung cancer in animal models, monitored tumor mutational burden in patients with colorectal cancer and stratified patients with glioblastoma. SCOPE can expedite readouts, augmenting the clinical use of EVs in precision oncology.

논문정보

형식Research article
게재일2024년 10월 (BRIC 등록일 2024-10-08)
연구진국내(교신)+국외 연구진
분야

댓글 작성 로그인

CV 열람하기

연락처 보기