CRISPR Gene Editing of Virulent Bacteriophage ICP1
- 28 April 2023
- journal article
- Published by Cold Spring Harbor Laboratory in Cold Spring Harbor Protocols
- Vol. 2024 (2)
- https://doi.org/10.1101/pdb.prot108316
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.This publication has 12 references indexed in Scilit:
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