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Issue 19, 2018
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A G-quadruplex motif at the 3′ end of sgRNAs improves CRISPR–Cas9 based genome editing efficiency

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Abstract

Originating as a component of prokaryotic adaptive immunity, the type II CRISPR/Cas9 system has been repurposed for targeted genome editing in various organisms. Although Cas9 can bind and cleave DNA efficiently under in vitro conditions, its activity inside a cell can vary dramatically between targets owing to the differences between genomic loci and the availability of enough Cas9/sgRNA (single guide RNA) complex molecules for cleavage. Most methods have so far relied on Cas9 protein engineering or base modifications in the sgRNA sequence to improve CRISPR/Cas9 activity. Here we demonstrate that a structure based rational design of sgRNAs can enhance the efficiency of Cas9 cleavage in vivo. By appending a naturally forming RNA G-quadruplex motif to the 3′ end of sgRNAs we can improve its stability and target cleavage efficiency in zebrafish embryos without inducing off-target activity, thereby underscoring its value in the design of better and optimized genome editing triggers.

Graphical abstract: A G-quadruplex motif at the 3′ end of sgRNAs improves CRISPR–Cas9 based genome editing efficiency

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Publication details

The article was received on 20 Nov 2017, accepted on 06 Feb 2018 and first published on 06 Feb 2018


Article type: Communication
DOI: 10.1039/C7CC08893K
Citation: Chem. Commun., 2018,54, 2377-2380
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    A G-quadruplex motif at the 3′ end of sgRNAs improves CRISPR–Cas9 based genome editing efficiency

    S. Nahar, P. Sehgal, M. Azhar, M. Rai, A. Singh, S. Sivasubbu, D. Chakraborty and S. Maiti, Chem. Commun., 2018, 54, 2377
    DOI: 10.1039/C7CC08893K

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