CRISPY-BRED and CRISPY-BRIP: efficient bacteriophage engineering

Abstract Genome engineering of bacteriophages provides opportunities for precise genetic dissection and for numerous phage applications including therapy. However, few methods are available for facile construction of unmarked precise deletions, insertions, gene replacements and point mutations in ba...

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Autores principales: Katherine S. Wetzel, Carlos A. Guerrero-Bustamante, Rebekah M. Dedrick, Ching-Chung Ko, Krista G. Freeman, Haley G. Aull, Ashley M. Divens, Jeremy M. Rock, Kira M. Zack, Graham F. Hatfull
Formato: article
Lenguaje:EN
Publicado: Nature Portfolio 2021
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Acceso en línea:https://doaj.org/article/b1b89e6a2de841798e39f54d739bc4c6
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Sumario:Abstract Genome engineering of bacteriophages provides opportunities for precise genetic dissection and for numerous phage applications including therapy. However, few methods are available for facile construction of unmarked precise deletions, insertions, gene replacements and point mutations in bacteriophages for most bacterial hosts. Here we describe CRISPY-BRED and CRISPY-BRIP, methods for efficient and precise engineering of phages in Mycobacterium species, with applicability to phages of a variety of other hosts. This recombineering approach uses phage-derived recombination proteins and Streptococcus thermophilus CRISPR-Cas9.