JACKIE: Fast Enumeration of Genome-Wide Single- and Multicopy CRISPR Target Sites and Their Off-Target Numbers.

JACKIE: Fast Enumeration of Genome-Wide Single- and Multicopy CRISPR Target Sites and Their Off-Target Numbers.
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DOI:
10.1089/crispr.2022.0042
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发表时间:
2022-08
期刊:
The CRISPR journal
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其他
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用于基因组和表观基因组编辑和成像的锌指蛋白、类转录激活子效应子和基于 CRISPR 的方法为研究基因组功能提供了强大的工具。靶向序列设计对于这些实验的成功至关重要。尽管现有的设计软件主要侧重于设计特定元素的目标序列,但我们在这里报告了 Jackie 和 Albert 的综合 K 聚体实例枚举器 (JACKIE) 的实现,这是一套用于枚举基因组中所有单拷贝和多拷贝位点的软件,可以将其纳入基因组规模设计,并与其他轨道一起加载到基因组浏览器上,以方便基于 Web 的图形用户界面设计。我们还实施快速算法来识别序列邻域或靶向序列的脱靶计数,以便可以在合理的时间内从数百万个设计序列中识别脱靶概率较低的设计。我们展示了 JACKIE 设计的 CRISPR 位点簇在基因组成像中的应用。
Zinc finger protein-, transcription activator like effector-, and CRISPR-based methods for genome and epigenome editing and imaging have provided powerful tools to investigate functions of genomes. Targeting sequence design is vital to the success of these experiments. Although existing design software mainly focus on designing target sequence for specific elements, we report here the implementation of Jackie and Albert's Comprehensive K-mer Instances Enumerator (JACKIE), a suite of software for enumerating all single- and multicopy sites in the genome that can be incorporated for genome-scale designs as well as loaded onto genome browsers alongside other tracks for convenient web-based graphic-user-interface-enabled design. We also implement fast algorithms to identify sequence neighborhoods or off-target counts of targeting sequences so that designs with low probability of off-target can be identified among millions of design sequences in reasonable time. We demonstrate the application of JACKIE-designed CRISPR site clusters for genome imaging.
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