한빛사 논문
Jeonghun Kwon1†, Minyoung Kim1†, Woochang Hwang2,3†, Anna Jo1, Gue‑Ho Hwang4, Minhee Jung1, Un Gi Kim1, Gang Cui5, Heonseok Kim6, Joon‑Ho Eom7, Junho K. Hur3,8, Junwon Lee5, Youngho Kim1, Jin‑soo Kim9, Sangsu Bae10 and Jungjoon K. Lee1*
1Toolgen, Seoul, Republic of Korea.
2Department of Pre‑Medicine, College of Medicine, Hanyang University, Seoul, Republic of Korea.
3Hanyang Institute of Bioscience and Biotechnology, Hanyang University, Seoul, Republic of Korea.
4Department of Chemistry, Hanyang University, Seoul, Republic of Korea.
5Department of Ophthalmology, Yonsei University College of Medicine, Seoul, Republic of Korea.
6Department of Medicine, Division of Oncology, Stanford University, Stanford, USA.
7National Institute of Food and Drug Safety Evaluation, Cheongju, Republic of Korea.
8Department of Genetics, College of Medicine, Hanyang University, Seoul, Republic of Korea.
9Center for Genome Engineering, Institute for Basic Science (IBS), Seoul, Republic of Korea.
10Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, Republic of Korea.
Corresponding author : Correspondence to Jungjoon K. Lee.
†Jeonghun Kwon, Minyoung Kim, and Woochang Hwang contributed equally to this work.
Abstract
We present a novel genome-wide off-target prediction method named Extru-seq and compare it with cell-based (GUIDE-seq), in vitro (Digenome-seq), and in silico methods using promiscuous guide RNAs with large numbers of valid off-target sites. Extru-seq demonstrates a high validation rate and retention of information about the intracellular environment, both beneficial characteristics of cell-based methods. Extru-seq also shows a low miss rate and could easily be performed in clinically relevant cell types with little optimization, which are major positive features of the in vitro methods. In summary, Extru-seq shows beneficial features of cell-based and in vitro methods.
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