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.
CITATION STYLE
Velusamy, T., Gowripalan, A., & Tscharke, D. C. (2020). CRISPR/Cas9-Based Genome Editing of HSV. In Methods in Molecular Biology (Vol. 2060, pp. 169–183). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9814-2_9
Mendeley helps you to discover research relevant for your work.