Efficient engineering of a bacteriophage genome using the type I-E CRISPR-Cas system.
RNA Biol
; 11(1): 42-4, 2014.
Article
em En
| MEDLINE
| ID: mdl-24457913
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.
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Base de dados:
MEDLINE
Assunto principal:
Bacteriófago T7
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Escherichia coli
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Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
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Sistemas CRISPR-Cas
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article