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- W2252568502 abstract "Genome-wide sgRNA libraries based on rules for on-target activity improve results of Cas9-based screens and facilitate a further refinement of on- and off-target prediction algorithms. CRISPR-Cas9–based genetic screens are a powerful new tool in biology. By simply altering the sequence of the single-guide RNA (sgRNA), one can reprogram Cas9 to target different sites in the genome with relative ease, but the on-target activity and off-target effects of individual sgRNAs can vary widely. Here, we use recently devised sgRNA design rules to create human and mouse genome-wide libraries, perform positive and negative selection screens and observe that the use of these rules produced improved results. Additionally, we profile the off-target activity of thousands of sgRNAs and develop a metric to predict off-target sites. We incorporate these findings from large-scale, empirical data to improve our computational design rules and create optimized sgRNA libraries that maximize on-target activity and minimize off-target effects to enable more effective and efficient genetic screens and genome engineering." @default.
- W2252568502 created "2016-06-24" @default.
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- W2252568502 date "2016-01-18" @default.
- W2252568502 modified "2023-10-18" @default.
- W2252568502 title "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9" @default.
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- W2252568502 doi "https://doi.org/10.1038/nbt.3437" @default.
- W2252568502 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4744125" @default.
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- W2252568502 hasPublicationYear "2016" @default.
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