FlashFry: a fast and flexible tool for large-scale CRISPR target design

Abstract

Abstract Background Genome-wide knockout studies, noncoding deletion scans, and other large-scale studies require a simple and lightweight framework that can quickly discover and score thousands of candidate CRISPR guides targeting an arbitrary DNA sequence. While several CRISPR web applications exist, there is a need for a high-throughput tool to rapidly discover and process hundreds of thousands of CRISPR targets. Results Here, we introduce FlashFry, a fast and flexible command-line tool for characterizing large numbers of CRISPR target sequences. With FlashFry, users can specify an unconstrained number of mismatches to putative off-targets, richly annotate discovered sites, and tag potential guides with commonly used on-target and off-target scoring metrics. FlashFry runs at speeds comparable to commonly used genome-wide sequence aligners, and output is provided as an easy-to-manipulate text file. Conclusions FlashFry is a fast and convenient command-line tool to discover and score CRISPR targets within large DNA sequences.

Keywords

• CRISPR
• Guide library
• Cas9
• Cpf1
• Off-target
• On-target