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Genetic Engineering Publications - GEG Tech top picks
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Genome-wide target specificities of CRISPR RNA-guided programmable deaminases - Nature Biotechnology
Base editors composed of Cas9 fused to a deaminase show high specificity in genome-wide analyses.
BigField GEG Tech's insight:
The off-target activity of CRISPR system is largely unknown. Here the scientists modify digested-genome sequencing (Digenome-seq) to assess the specificity of a programmable deaminase composed of a Cas9 nickase (nCas9) and the deaminase APOBEC1 in the human genome. Digenome-seq is sensitive enough to capture off-target sites with a substitution frequency of 0.1%. Notably, off-target sites of the base editors are often different from those of Cas9 alone, calling for independent assessment of their genome-wide specificities. 
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The scientists recently reported Digenome-seq (digested genome sequencing), a method for in vitro identification of potential off-target sites, and They evaluated the specificity of CRISPR–Cas9 and CRISPR–Cpf1 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 their previous report required extensive manual interaction and produced several large intermediate files, resulting in a long running time. Here, the scientists 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 their 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.