The DOI matches its Crossref record; 40 of the 40 references it cites were found in a registry or an index.
sgRNAcas9: A Software Package for Designing CRISPR sgRNA and Evaluating Potential Off-Target Cleavage Sites
Open accessAbstract
Although the CRISPR/Cas9/sgRNA system efficiently cleaves intracellular DNA at desired target sites, major concerns remain on potential "off-target" cleavage that may occur throughout the whole genome. In order to improve CRISPR-Cas9 specificity for targeted genome editing and transcriptional control, we describe a bioinformatics tool "sgRNAcas9", which is a software package developed for fast design of CRISPR sgRNA with minimized off-target effects. This package consists of programs to perform a search for CRISPR target sites (protospacers) with user-defined parameters, predict genome-wide Cas9 potential off-target cleavage sites (POT), classify the POT into three categories, batch-design oligonucleotides for constructing 20-nt (nucleotides) or truncated sgRNA expression vectors, extract desired length nucleotide sequences flanking the on- or off-target cleavage sites for designing PCR primer pairs to validate the mutations by T7E1 cleavage assay. Importantly, by identifying potential off-target sites in silico, the sgRNAcas9 allows the selection of more specific target sites and aids the identification of bona fide off-target sites, significantly facilitating the design of sgRNA for genome editing applications. sgRNAcas9 software package is publicly available at BiooTools website (www.biootools.com) under the terms of the GNU General Public License.
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Cites
Multiplex Genome Engineering Using CRISPR/Cas Systems
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RNA-Guided Human Genome Engineering via Cas9
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CRISPR Provides Acquired Resistance Against Viruses in Prokaryotes
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Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression
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Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity
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Self-citationHigh-Frequency off-Target Mutagenesis Induced by CRISPR-Cas Nucleases in Human Cells
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“Previous reports have shown that Cas9 nuclease cuts 3-nt upstream of the PAM site [34].”
BackgroundInfluentialGenetic screens in human cells using the CRISPR-Cas9 system
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RNA-guided editing of bacterial genomes using CRISPR-Cas systems
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Cited by
The new frontier of genome engineering with CRISPR-Cas9
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CRISPR-Cas9 Based Engineering of Actinomycetal Genomes
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Insert, remove or replace: A highly advanced genome editing system using CRISPR/Cas9
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Metadata
- Authors
- Xie, S., Shen, B., Zhang, C., Huang, X., & Zhang, Y.
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2014-06-23
- Volume
- 9
- Issue
- 6
- Pages
- e100448
- ISSN
- 1932-6203 (ISSN-L)
- OpenAlex
- W2133836136
- Cited by
- 417
- References
- 40
- FWCI
- 11.32
- Type
- journal article
- Language
- en
- Open access
- gold
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