0000000001162410
AUTHOR
Paul R. Macdonald
Comparison of CRISPR and marker based methods for the engineering of phage T7
With the recent rise in interest in using lytic bacteriophages as therapeutic agents, there is an urgent requirement to understand their fundamental biology to enable the engineering of their genomes. Current methods of phage engineering rely on homologous recombination, followed by a system of selection to identify recombinant phages. For bacteriophage T7, the host genescmkortrxhave been used as a selection mechanism along with both type I and II CRISPR systems to select against wild-type phage and enrich for the desired mutant. Here we systematically compare all three systems; we show that the use of marker-based selection is the most efficient method and we use this to generate multiple …
Comparison of CRISPR and Marker-Based Methods for the Engineering of Phage T7
This article belongs to the Section Bacterial Viruses.