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. 2020 Jun 13;5(2):99-102.
doi: 10.1016/j.synbio.2020.05.005. eCollection 2020 Jun.

Designing sgRNAs for CRISPR-BEST base editing applications with CRISPy-web 2.0

Kai Blin  1 , Simon Shaw  1 , Yaojun Tong  1 , Tilmann Weber  1
Affiliations

Designing sgRNAs for CRISPR-BEST base editing applications with CRISPy-web 2.0

Kai Blin et al. Synth Syst Biotechnol. .

Erratum in

  • Erratum regarding previously published articles.
    [No authors listed] [No authors listed] Synth Syst Biotechnol. 2020 Oct 14;5(4):330-331. doi: 10.1016/j.synbio.2020.10.001. eCollection 2020 Dec. Synth Syst Biotechnol. 2020. PMID: 33102827 Free PMC article.

Abstract

CRISPR/Cas9 systems are an established tool in genome engineering. As double strand breaks caused by the standard Cas9-based knock-out techniques can be problematic in some organisms, new systems were developed that can efficiently create knock-outs without causing double strand breaks to elegantly sidestep these issues. The recently published CRISPR-BEST base editor system for actinobacteria is built around a C to T or A to G base exchange. These base editing systems however require additional constraints to be considered for designing the sgRNAs. Here, we present an updated version of the interactive CRISPy-web single guide RNA design tool https://crispy.secondarymetabolites.org/that was built to support "classical" CRISPR and now also CRISPR-BEST workflows.

Keywords: Base editor; CRISPR; CRISPR-BEST; Genome editing; Webserver; sgRNA.

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Figures

Fig. 1
Fig. 1
A: Target selection dialog with unfolded “Expert settings” to select the Cas enzyme (PAM and spacer size are filled out automatically) and base editor properties, and unfolded “Usage hints” describing the syntax to define the target sequence. B: Display of CRISPy-web sgRNA predictions for the actinorhodin-BGC of S. coelicolor. C: Selection buttons to activate base editor/CRISPR-BEST mode. D: sgRNA predictions for CRISPR-cBEST base editing applications. Only sgRNAs are displayed that will lead to STOP mutations in the selected target gene (indicated by *); if there are further C bases within the editing window, all potential amino acid exchanges are displayed.
See this image and copyright information in PMC

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