Genetic Engineering of Therapeutic Phages Using Type III CRISPR-Cas Systems

Courtney M. Hill, Asma Hatoum-Aslan

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The functional characterization of “hypothetical” phage genes is a major bottleneck in basic and applied phage research. To compound this issue, the most suitable phages for therapeutic applications—the strictly lytic variety—are largely recalcitrant to classical genetic techniques due to low recombination rates and lack of selectable markers. Here we describe methods for fast and effective phage engineering that rely upon a Type III-A CRISPR-Cas system. In these methods, the CRISPR-Cas system is used as a powerful counterselection tool to isolate rare phage recombinants.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages279-299
Number of pages21
DOIs
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
Volume2734
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Bacteriophage engineering
  • CRISPR-Cas10
  • Phage genome editing
  • Staphylococci
  • Type III-A CRISPR-Cas

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

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