Lambda as a Cloning Vector

Lech, Karen; Brent, Roger; Irwin, Nina

doi:10.1002/0471142727.mb0110s00

Cited by 2 publications

(1 citation statement)

References 11 publications

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“…The PCR product was introduced by homologous recombination into a λ prophage (Fig. 1), in place of genes that were shown to be dispensable for phage growth and lysogenization (20,21). A CRISPR array, encoding spacers that target conserved sequences of the resistance genes ndm-1 and ctx-M-15, was also introduced into the same lysogen, immediately downstream of the cas genes ( Fig.…”

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confidence: 99%

Temperate and lytic bacteriophages programmed to sensitize and kill antibiotic-resistant bacteria

Yosef

Manor

Kiro

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

401

284

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The increasing threat of pathogen resistance to antibiotics requires the development of novel antimicrobial strategies. Here we present a proof of concept for a genetic strategy that aims to sensitize bacteria to antibiotics and selectively kill antibiotic-resistant bacteria. We use temperate phages to deliver a functional clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPRassociated (Cas) system into the genome of antibiotic-resistant bacteria. The delivered CRISPR-Cas system destroys both antibiotic resistance-conferring plasmids and genetically modified lytic phages. This linkage between antibiotic sensitization and protection from lytic phages is a key feature of the strategy. It allows programming of lytic phages to kill only antibiotic-resistant bacteria while protecting antibiotic-sensitized bacteria. Phages designed according to this strategy may be used on hospital surfaces and hand sanitizers to facilitate replacement of antibioticresistant pathogens with sensitive ones.CRISPR-Cas | positive selection | lysogenization | ex vivo treatment

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confidence: 99%