CRISPR/Cas9-Based Genome Editing of HSV

Thilaga Velusamy, Anjali Gowripalan, David C. Tscharke*

*Corresponding author for this work

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

    12 Citations (Scopus)

    Abstract

    The CRISPR/Cas9 gene editing system is a robust and versatile technology that has revolutionized our capacity for genome engineering and is applicable in a wide range of organisms, including large dsDNA viruses. Here we provide an efficient methodology that can be used both for marker-based and for marker-free CRISPR/Cas9-mediated editing of the HSV-1 genome. In our method, Cas9, guide RNAs and a homology-directed repair template are provided to cells by cotransection of plasmids, followed by introduction of the HSV genome by infection. This method offers a great deal of flexibility, facilitating editing of the HSV genome that spans the range from individual nucleotide changes to large deletions and insertions.

    Original languageEnglish
    Title of host publicationMethods in Molecular Biology
    PublisherHumana Press Inc.
    Pages169-183
    Number of pages15
    DOIs
    Publication statusPublished - 2020

    Publication series

    NameMethods in Molecular Biology
    Volume2060
    ISSN (Print)1064-3745
    ISSN (Electronic)1940-6029

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