ge-CRISPR - An integrated pipeline for the prediction and analysis of sgRNAs genome editing efficiency for CRISPR/Cas system

被引:35
作者
Kaur, Karambir [1 ]
Gupta, Amit Kumar [1 ]
Rajput, Akanksha [1 ]
Kumar, Manoj [1 ]
机构
[1] CSIR, Inst Microbial Technol, Bioinformat Ctr, Sect 39A, Chandigarh 160036, India
关键词
ADAPTIVE IMMUNITY; VERSATILE TOOL; RNA; DNA; CAS9; SPECIFICITY; MUTATIONS; SOFTWARE; SEQUENCE; PLANTS;
D O I
10.1038/srep30870
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Genome editing by sgRNA a component of CRISPR/Cas system emerged as a preferred technology for genome editing in recent years. However, activity and stability of sgRNA in genome targeting is greatly influenced by its sequence features. In this endeavor, a few prediction tools have been developed to design effective sgRNAs but these methods have their own limitations. Therefore, we have developed "ge-CRISPR" using high throughput data for the prediction and analysis of sgRNAs genome editing efficiency. Predictive models were employed using SVM for developing pipeline-1 (classification) and pipeline-2 (regression) using 2090 and 4139 experimentally verified sgRNAs respectively from Homo sapiens, Mus musculus, Danio rerio and Xenopus tropicalis. During 10-fold cross validation we have achieved accuracy and Matthew's correlation coefficient of 87.70% and 0.75 for pipeline-1 on training dataset (T-1840) while it performed equally well on independent dataset (V-250). In pipeline-2 we attained Pearson correlation coefficient of 0.68 and 0.69 using best models on training (T-3169) and independent dataset (V-520) correspondingly. ge-CRISPR (http://bioinfo.imtech.res.in/manojk/gecrispr/) for a given genomic region will identify potent sgRNAs, their qualitative as well as quantitative efficiencies along with potential off-targets. It will be useful to scientific community engaged in CRISPR research and therapeutics development.
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页数:12
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