Editing of Phage Genomes - Recombineering-Assisted SpCas9 Modification of Model Coliphages T7, T5, and T3

TY - JOUR

T1 - Editing of Phage Genomes - Recombineering-Assisted SpCas9 Modification of Model Coliphages T7, T5, and T3

AU - Isaev, A.

AU - Andriianov, A.

AU - Znobishcheva, E.

AU - Zorin, E.

AU - Morozova, N.

AU - Severinov, K.

PY - 2022/11/1

Y1 - 2022/11/1

N2 - Bacteriophages-viruses that infect bacterial cells - are the most abundant biological entities on Earth. The use of phages in fundamental research and industry requires tools for precise manipulation of their genomes. Yet, compared to bacterial genome engineering, modification of phage genomes is challenging because of the lack of selective markers and thus requires laborious screenings of recombinant/mutated phage variants. The development of the CRISPR-Cas technologies allowed to solve this issue by the implementation of negative selection that eliminates the parental phage genomes. In this manuscript, we summarize current methods of phage genome engineering and their coupling with CRISPR-Cas technologies. We also provide examples of our successful application of these methods for introduction of specific insertions, deletions, and point mutations in the genomes of model Escherichia coli lytic phages T7, T5, and T3.

AB - Bacteriophages-viruses that infect bacterial cells - are the most abundant biological entities on Earth. The use of phages in fundamental research and industry requires tools for precise manipulation of their genomes. Yet, compared to bacterial genome engineering, modification of phage genomes is challenging because of the lack of selective markers and thus requires laborious screenings of recombinant/mutated phage variants. The development of the CRISPR-Cas technologies allowed to solve this issue by the implementation of negative selection that eliminates the parental phage genomes. In this manuscript, we summarize current methods of phage genome engineering and their coupling with CRISPR-Cas technologies. We also provide examples of our successful application of these methods for introduction of specific insertions, deletions, and point mutations in the genomes of model Escherichia coli lytic phages T7, T5, and T3.

KW - CRISPR-Cas

KW - T3

KW - T5

KW - T7

KW - bacteriophage

KW - genome editing

KW - homologous recombination

KW - phage rebooting

KW - phage therapy

KW - recombineering

UR - https://www.scopus.com/pages/publications/85143563977

UR - https://www.scopus.com/pages/publications/85143563977#tab=citedBy

U2 - 10.31857/S002689842206009X

DO - 10.31857/S002689842206009X

M3 - Article

C2 - 36475474

SN - 0026-8984

VL - 56

SP - 883

JO - Molekuliarnaia biologiia

JF - Molekuliarnaia biologiia

IS - 6

ER -