Detection of unamplified target genes via CRISPR–Cas9 immobilized on a graphene field-effect transistor View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-03-25

AUTHORS

Reza Hajian, Sarah Balderston, Thanhtra Tran, Tara deBoer, Jessy Etienne, Mandeep Sandhu, Noreen A. Wauford, Jing-Yi Chung, Jolie Nokes, Mitre Athaiya, Jacobo Paredes, Regis Peytavi, Brett Goldsmith, Niren Murthy, Irina M. Conboy, Kiana Aran

ABSTRACT

Most methods for the detection of nucleic acids require many reagents and expensive and bulky instrumentation. Here, we report the development and testing of a graphene-based field-effect transistor that uses clustered regularly interspaced short palindromic repeats (CRISPR) technology to enable the digital detection of a target sequence within intact genomic material. Termed CRISPR–Chip, the biosensor uses the gene-targeting capacity of catalytically deactivated CRISPR-associated protein 9 (Cas9) complexed with a specific single-guide RNA and immobilized on the transistor to yield a label-free nucleic-acid-testing device whose output signal can be measured with a simple handheld reader. We used CRISPR–Chip to analyse DNA samples collected from HEK293T cell lines expressing blue fluorescent protein, and clinical samples of DNA with two distinct mutations at exons commonly deleted in individuals with Duchenne muscular dystrophy. In the presence of genomic DNA containing the target gene, CRISPR–Chip generates, within 15 min, with a sensitivity of 1.7 fM and without the need for amplification, a significant enhancement in output signal relative to samples lacking the target sequence. CRISPR–Chip expands the applications of CRISPR–Cas9 technology to the on-chip electrical detection of nucleic acids. An electrical biosensor combining CRISPR–Cas9 and a graphene field-effect transistor detects target genes in purified genomic samples at high sensitivity, within 15 minutes, and without the need for amplification. More... »

PAGES

1-11

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/s41551-019-0371-x

    DOI

    http://dx.doi.org/10.1038/s41551-019-0371-x

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1112971580


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