Abstract：
:A major limitation hindering the widespread use of synthetic phages in medical and industrial settings is the lack of an efficient phage-engineering platform. Classical T4 phage engineering and several newly proposed methods are often inefficient and time consuming and consequently, only able to produce an inconsistent range of genomic editing rates between 0.03-3%. Here, we review and present new understandings of the CRISPR/Cas9 assisted genome engineering technique that significantly improves the genomic editing rate of T4 phages. Our results indicate that crRNAs selection is a major rate limiting factor in T4 phage engineering via CRISPR/Cas9. We were able to achieve an editing rate of > 99% for multiple genes that functionalizes the phages for further applications. We envision that this improved phage-engineering platform will accelerate the fields of individualized phage therapy, biocontrol, and rapid diagnostics.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Duong MM,Carmody CM,Ma Q,Peters JE,Nugen SRdoi
10.1038/s41598-020-75426-6subject
Has Abstractpub_date
2020-10-26 00:00:00pages
18229issue
1issn
2045-2322pii
10.1038/s41598-020-75426-6journal_volume
10pub_type
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