CRISPR Gene Editing of a Virulent Bacteriophage

Genome editing by site-directed mutagenesis is an important tool in biological research. CRISPR gene editing is the latest such tool developed, and one that is widely applicable to study organisms from all kingdoms of life. Here, I introduce a method for making site-directed, defined mutations in a virulent bacteriophage (a bacterial virus) using CRISPR gene editing. The ability to precisely edit the genomes of virulent phages will facilitate the study of their gene requirements for infection of host bacteria and advance our ability to engineer phages for use as therapeutic agents to combat bacterial infections. The protocol introduced here was developed as part of Cold Spring Harbor's Advanced Bacterial Genetics course.