Tongji University Cancer Center, Shanghai Tenth People's Hospital, School of Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
School of Medicine, Tongji University, Shanghai 200092, China.
Brief Bioinform. 2022 Jan 17;23(1). doi: 10.1093/bib/bbab505.
Clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR/Cas9) technology has become a popular tool for the study of genome function, and the use of this technology can achieve large-scale screening studies of specific phenotypes. Several analysis tools for CRISPR/Cas9 screening data have been developed, while high false positive rate remains a great challenge. To this end, we developed iCRISEE, an integrative analysis of CRISPR ScrEEn by reducing false positive hits. iCRISEE can dramatically reduce false positive hits and it is robust to different single guide RNA (sgRNA) library by introducing precise data filter and normalization, model selection and valid sgRNA number correction in data preprocessing, sgRNA ranking and gene ranking. Furthermore, a powerful web server has been presented to automatically complete the whole CRISPR/Cas9 screening analysis, where we integrated the main hypothesis in multiple algorithms as a full workflow, including quality control, sgRNA extracting, sgRNA alignment, sgRNA ranking, gene ranking and pathway enrichment. In addition, output of iCRISEE, including result mapping, sample clustering, sgRNA ranking and gene ranking, can be easily visualized and downloaded for publication. Taking together, iCRISEE presents to be the state-of-the-art and user-friendly tool for CRISPR screening data analysis. iCRISEE is available at https://www.icrisee.com.
CRISPR/Cas9 技术已成为研究基因组功能的热门工具,利用该技术可以实现特定表型的大规模筛选研究。已经开发了几种用于 CRISPR/Cas9 筛选数据的分析工具,而高假阳性率仍然是一个巨大的挑战。为此,我们开发了 iCRISEE,一种通过减少假阳性命中来整合 CRISPR 筛选的分析。iCRISEE 可以显著减少假阳性命中,并且通过在数据预处理中引入精确的数据过滤和归一化、模型选择和有效 sgRNA 数量校正、sgRNA 排名和基因排名,它对不同的单指导 RNA(sgRNA)文库具有很强的稳健性。此外,我们还提供了一个强大的网络服务器,用于自动完成整个 CRISPR/Cas9 筛选分析,其中我们将多个算法中的主要假设集成到一个完整的工作流程中,包括质量控制、sgRNA 提取、sgRNA 比对、sgRNA 排名、基因排名和途径富集。此外,iCRISEE 的输出,包括结果映射、样本聚类、sgRNA 排名和基因排名,可以轻松可视化和下载以供发布。总之,iCRISEE 是一种用于 CRISPR 筛选数据分析的最先进且用户友好的工具。iCRISEE 可在 https://www.icrisee.com 上获得。