Efficient engineering of a bacteriophage genome using the type I-E CRISPR-Cas system

Ruth Kiro, Dror Shitrit, Udi Qimron*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

116 Scopus citations

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) system has recently been used to engineer genomes of various organisms, but surprisingly, not those of bacteriophages (phages). Here we present a method to genetically engineer the Escherichia coli phage T7 using the type I-E CRISPR-Cas system. T7 phage genome is edited by homologous recombination with a DNA sequence flanked by sequences homologous to the desired location. Non-edited genomes are targeted by the CRISPR-Cas system, thus enabling isolation of the desired recombinant phages. This method broadens CRISPR Cas-based editing to phages and uses a CRISPR-Cas type other than type II. The method may be adjusted to genetically engineer any bacteriophage genome.

Original languageEnglish
Pages (from-to)42-44
Number of pages3
JournalRNA Biology
Volume11
Issue number1
DOIs
StatePublished - Jan 2014

Funding

FundersFunder number
FP7/207
Seventh Framework Programme336079
European Research Council

    Keywords

    • Bacteriophage T7
    • Escherichia coli
    • Genetic engineering
    • Homologous recombination
    • Negative selection
    • Positive selection
    • Spacer targeting

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