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AsCRISPR:精准医学中用于等位基因特异性单指导 RNA 设计的网络服务器。

AsCRISPR: A Web Server for Allele-Specific Single Guide RNA Design in Precision Medicine.

机构信息

National Clinical Research Center for Geriatric Disorders, Department of Geriatrics, Xiangya Hospital, Central South University, Changsha, P.R. China.

出版信息

CRISPR J. 2020 Dec;3(6):512-522. doi: 10.1089/crispr.2020.0071.

Abstract

Allele-specific genomic targeting by CRISPR is a versatile strategy that has been increasingly exploited not only in treating inherited dominant diseases and mutation-driven cancers, but also in other important fields such as genome imprinting, haploinsufficiency, and genome loci imaging. Despite its tremendous utilities, few bioinformatic tools have been implemented for the allele-specific purpose of CRISPR. We thus developed AsCRISPR (llele-pecific CRISPR), a comprehensive web tool to aid the design of short-guide RNA (sgRNA) sequences that can discriminate between alleles. AsCRISPR allows users to analyze both their own identified variants and heterozygous single nucleotide polymorphisms and, importantly, output the candidate sgRNAs and their quality control information. To facilitate targeting dominant diseases, AsCRISPR analyzed dominant single nucleotide variants (SNVs) retrieved from ClinVar and OMIM databases, and generated a dominant database of candidate-discriminating sgRNAs that may specifically target the alternative allele for each dominant SNV site. Moreover, a validated database was established, which manually curated the discriminating sgRNAs that were experimentally validated in the mounting literature for multiple allele-specific purposes.

摘要

通过 CRISPR 进行等位基因特异性基因组靶向是一种多功能策略,不仅在治疗遗传显性疾病和突变驱动的癌症方面得到了越来越多的应用,而且在基因组印迹、单倍不足和基因组位点成像等其他重要领域也得到了应用。尽管 CRISPR 具有巨大的实用价值,但针对等位基因特异性的生物信息学工具却很少。因此,我们开发了 AsCRISPR(allele-pecific CRISPR),这是一个全面的网络工具,用于辅助设计能够区分等位基因的短向导 RNA(sgRNA)序列。AsCRISPR 允许用户分析他们自己识别的变体和杂合单核苷酸多态性,并且重要的是,输出候选 sgRNA 及其质量控制信息。为了便于靶向显性疾病,AsCRISPR 分析了从 ClinVar 和 OMIM 数据库中检索到的显性单核苷酸变异(SNV),并生成了候选区分 sgRNA 的显性数据库,这些 sgRNA 可能专门针对每个显性 SNV 位点的替代等位基因。此外,还建立了一个经过验证的数据库,该数据库手动整理了在不断增加的文献中针对多种等位基因特异性目的进行实验验证的区分 sgRNA。

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