JMB-HEADER RAS-JOURNALS EIMB Pleiades Publishing

RUS

             

ENG

YearIMPACT-FACTOR
2022  1,200
2021  1,540
2020  1,374
2019  1,023
2018  0,932
2017  0,977
2016  0,799
2015  0,662
2014  0,740
2013  0,739
2012  0,637
2011  0,658
2010  0,654
2009  0,570
2008  0,849
2007  0,805
2006  0,330
2005  0,435
2004  0,623
2003  0,567
2002  0,641
2001  0,490
2000  0,477
1999  0,762
1998  0,785
1997  0,507
1996  0,518
1995  0,502
Vol 56(2022) N 6 p. 801-815; DOI 10.1134/S0026893322060073 Full Text

A. Isaev1*, A. Andriianov1, E. Znobishcheva2, E. Zorin1, N. Morozova2, K. Severinov1,2,3**

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

1Skolkovo Institute of Science and Technology, Skolkovo, Moscow, 143028 Russia
2Peter the Great St Petersburg State Polytechnic University, St Petersburg, 195251 Russia
3Waksman Institute of Microbiology, Piscataway, NJ 08854 USA

*tcft18@gmail.com
**severik@waksman.rutgers.edu
Received - 2022-04-01; Revised - 2022-05-22; Accepted - 2022-05-22

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.

bacteriophage, CRISPR-Cas, genome editing, recombineering, phage therapy, homologous recombination, phage rebooting, T7, T5, T3



JMB-FOOTER RAS-JOURNALS