CRISPR Gene Editing of Virulent Bacteriophage ICP1
- Department of Molecular Biology and Microbiology, Tufts University, School of Medicine, Boston, Massachusetts 02067, USA
- ↵1Correspondence: andrew.camilli{at}tufts.edu
Abstract
Tools for site-directed mutagenesis of virulent bacteriophages (phages; viruses of bacteria) have traditionally lagged those for bacteria, hindering their study. CRISPR gene editing represents a new and highly efficient method for editing virulent phage genomes. Here, I describe methods for using CRISPR gene editing for site-directed mutagenesis of ICP1, a virulent phage of Vibrio cholerae. The first section outlines methods of constructing a plasmid for CRISPR editing of an ICP1 gene. The second section outlines methods of transferring the plasmid to an editing-competent strain of V. cholerae. The third section outlines methods of selecting for and storing the edited phage.
Footnotes
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From the Experiments in Bacterial Genetics collection, edited by Lionello Bossi, Andrew Camilli, and Angelika Gründling.
