1Division of Theoretical Bioinformatics, German Cancer Research Center (DKFZ), Heidelberg, Germany. 2Center for Genome Engineering, Institute for Basic Science, Seoul, South Korea. 3Department of Chemistry, Hanyang University, Seoul, South Korea. 4Department of Chemistry, Seoul National University, Seoul, South Korea. 5Institute of Nano Science and Technology, Hanyang University, Seoul, South Korea.
*Correspondence should be addressed to S.B. or J.-S.K..
Abstract
To the Editor: We recently reported Digenome-seq (digested genome sequencing), a method for in vitro identification of potential off-target sites, and we evaluated the specificity of CRISPR–Cas9 (refs. 1,2) and CRISPR–Cpf1 (ref. 3) endonuclease by whole-genome sequencing. Digenome-seq pinpoints the exact location of double-strand break (DSB) sites by recognizing specific patterns of aligned reads. However, the analysis pipeline presented in our previous report required extensive manual interaction and produced several large intermediate files, resulting in a long running time. Here, we present a redesigned analysis tool for Digenome-seq data that runs on web browsers. The core algorithm of the tool is written in C++ and compiled to asm.js (http://asmjs.org/), a preoptimized subset of JavaScript. Users can instantly perform the complete analysis in an ordinary web browser (Supplementary Note 1) with fast execution speed without uploading any data to a server and without local tool installation. In our benchmark, the full analysis for 100 GB of BAM file took 3 h for whole analysis on Intel i5 3570k central processing unit in a single thread.
1. 논문관련 분야의 소개, 동향, 전망을 설명, 연구과정에서 생긴 에피소드
CRISPR-Cas 시스템을 이용한 유전체 교정 (genome editing) 기술은 2012년 처음 CRISPR-Cas9을 이용한 유전체 교정이 가능하다는 보고가 나온 이래로 불과 5년 사이 눈부신 발전을 거듭해 왔습니다. 이러한 CRISPR-Cas nuclease 들은 1) target cleavage site를 guide RNA-DNA hybridization을 통해 찾아내기...