HiTSelect: a comprehensive tool for high-complexity-pooled screen analysis.

Authors: Diaz AA, Qin H, Ramalho-Santos M, Song JS.

Reference: Nucleic Acids Res. 2015 Feb 18;43(3):e16. doi: 10.1093/nar/gku1197. Epub 2014 Nov 26.

PMID: 25428347

Abstract: Genetic screens of an unprecedented scale have recently been made possible by the availability of high-complexity libraries of synthetic oligonucleotides designed to mediate either gene knockdown or gene knockout, coupled with next-generation sequencing. However, several sources of random noise and statistical biases complicate the interpretation of the resulting high-throughput data. We developed HiTSelect, a comprehensive analysis pipeline for rigorously selecting screen hits and identifying functionally relevant genes and pathways by addressing off-target effects, controlling for variance in both gene silencing efficiency and sequencing depth of coverage and integrating relevant metadata. We document the superior performance of HiTSelect using data from both genome-wide RNAi and CRISPR/Cas9 screens. HiTSelect is implemented as an open-source package, with a user-friendly interface for data visualization and pathway exploration. Binary executables are available at sourceforge.net/projects/hitselect/, and the source code is available at github.com/diazlab/HiTSelect.

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