Abstract
The CRISPR/Cas9 system has been developed as a powerful technology for both targeted genome editing and gene regulation. However, the design of efficient single-guide RNAs (sgRNAs) remains challenging with the consideration of many criteria. In this section, we introduce how to design sgRNA sequences and build genome-wide sgRNA library using CRISPR-ERA, which is one of the state-of-the-art designer webserver tools for sgRNA design based on a set of sgRNA design rules summarized from published reports.
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References
Qi LS, Larson MH, Gilbert LA et al (2013) Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell 152:1173–1183
Nakamura M, Srinivasan P, Chavez M et al (2019) Anti-crispr-mediated control of gene editing and synthetic circuits in eukaryotic cells. Nat Commun 10:194–204
Liu H, Wei Z, Dominguez A et al (2015) CRISPR-ERA: a comprehensive design tool for CRISPR-mediated gene editing, repression and activation. Bioinformatics 31:3676–3678
Liu H, Wang X, Qi LS (2017) Using CRISPR-ERA webserver for sgRNA design. Bio-protocol 7:e2522
Doudna JA, Charpentier E (2014) Genome editing. The new frontier of genome engineering with CRISPR-Cas9. Science 346:1258096
La Russa MF, Qi LS (2015) The new state of the art: Cas9 for gene activation and repression. Mol Cell Biol 35:3800–3809
Liu Y, Yu C, Daley TP et al (2018) Crispr activation screens systematically identify factors that drive neuronal fate and reprogramming. Cell Stem Cell 23:758–771.e8
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Liu, H., Wang, X. (2021). CRISPR-ERA: A Webserver for Guide RNA Design of Gene Editing and Regulation. In: Marchisio, M.A. (eds) Computational Methods in Synthetic Biology. Methods in Molecular Biology, vol 2189. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0822-7_5
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DOI: https://doi.org/10.1007/978-1-0716-0822-7_5
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0821-0
Online ISBN: 978-1-0716-0822-7
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