Benchmark Software and Data for Evaluating CRISPR-Cas9 Experimental Pipelines Through the Assessment of a Calibration Screen.

IF 3.7 4区 生物学 Q2 GENETICS & HEREDITY CRISPR Journal Pub Date : 2024-01-02 DOI:10.1089/crispr.2023.0040
Raffaele M Iannuzzi, Ichcha Manipur, Clare Pacini, Fiona M Behan, Mario R Guarracino, Mathew J Garnett, Aurora Savino, Francesco Iorio
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Abstract

Genome-wide genetic screens using CRISPR-guide RNA libraries are widely performed in mammalian cells to functionally characterize individual genes and for the discovery of new anticancer therapeutic targets. As the effectiveness of such powerful and precise tools for cancer pharmacogenomics is emerging, tools and methods for their quality assessment are becoming increasingly necessary. Here, we provide an R package and a high-quality reference data set for the assessment of novel experimental pipelines through which a single calibration experiment has been executed: a screen of the HT-29 human colorectal cancer cell line with a commercially available genome-wide library of single-guide RNAs. This package and data allow experimental researchers to benchmark their screens and produce a quality-control report, encompassing several quality and validation metrics. The R code used for processing the reference data set, for its quality assessment, as well as to evaluate the quality of a user-provided screen, and to reproduce the figures presented in this article is available at https://github.com/DepMap-Analytics/HT29benchmark. The reference data is publicly available on FigShare.

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通过校准筛选评估 CRISPR-Cas9 实验流程的基准软件和数据。
利用 CRISPR 引导的 RNA 文库在哺乳动物细胞中广泛开展全基因组遗传筛选,以确定单个基因的功能特征和发现新的抗癌治疗靶点。随着这种强大而精确的癌症药物基因组学工具的有效性不断显现,对其质量进行评估的工具和方法也变得越来越必要。在此,我们提供了一个 R 软件包和一个高质量的参考数据集,用于评估新的实验管道,并通过该管道执行了一次校准实验:使用市售的全基因组单导 RNA 文库对 HT-29 人类结直肠癌细胞系进行筛选。实验研究人员可以利用这个软件包和数据对他们的筛选进行基准测试,并生成一份包含多个质量和验证指标的质量控制报告。用于处理参考数据集、进行质量评估、评估用户提供的筛选质量以及复制本文所展示图表的 R 代码可在 https://github.com/DepMap-Analytics/HT29benchmark 网站上获取。参考数据可在 FigShare 上公开获取。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CRISPR Journal
CRISPR Journal Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.30
自引率
2.70%
发文量
76
期刊介绍: In recognition of this extraordinary scientific and technological era, Mary Ann Liebert, Inc., publishers recently announced the creation of The CRISPR Journal -- an international, multidisciplinary peer-reviewed journal publishing outstanding research on the myriad applications and underlying technology of CRISPR. Debuting in 2018, The CRISPR Journal will be published online and in print with flexible open access options, providing a high-profile venue for groundbreaking research, as well as lively and provocative commentary, analysis, and debate. The CRISPR Journal adds an exciting and dynamic component to the Mary Ann Liebert, Inc. portfolio, which includes GEN (Genetic Engineering & Biotechnology News) and more than 80 leading peer-reviewed journals.
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