Overview of guide RNA design tools for CRISPR-Cas9 genome editing technology
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CRISPR-Cas (Clustered, Regularly Interspaced, Short Palindromic Repeats–CRISPR-associated (Cas)) RNA guided endonuclease has emerged as the most effective and widely used genome editing technology, which has become the most exciting and rapidly advancing research field. Efficient genome editing by the CRISPR-Cas9 system has been demonstrated in many species, and several laboratories have established CRISPR-Cas9 as a screening tool for systematic genetic analysis, similar to shRNA screening. At least three companies have been founded to leverage this technology for therapeutic uses. To facilitate the implementation of this technology, many software tools have been developed to identify guide RNAs that effectively target a desired genomic region. Here, I provide an overview of the technology, focusing on guide RNA design principles, available software tools and their strengths and weaknesses.
KeywordsCRISPR-Cas9 genome editing gRNA design off-target analysis gRNA efficacy
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- Enuameh M S, Asriyan Y, Richards A, Christensen R G, Hall V L, Kazemian M, Zhu C, Pham H, Cheng Q, Blatti C, Brasefield J A, Basciotta M D, Ou J, McNulty J C, Zhu L J, Celniker S E, Sinha S, Stormo G D, Brodsky M H, Wolfe S A (2013). Global analysis of Drosophila Cys2-His2 zinc finger proteins reveals a multitude of novel recognition motifs and binding determinants. Genome Res, 23 (6): 928–940PubMedCentralCrossRefPubMedGoogle Scholar
- Xiao A, Cheng Z, Kong L, Zhu Z, Lin S, Gao G, Zhang B (2014). CasOT: a genome-wide Cas9/gRNA off-target searching tool. BioinformaticsGoogle Scholar
- Xu H, Xiao T, Chen C H, Li W, Meyer C, Wu Q, Wu D, Cong L, Zhang F, Liu J S, Brown M, Liu S X (2015). Sequence determinants of improved CRISPR sgRNA design. Genome Res: gr.191452.115Google Scholar