Abstract: Embodiments of the invention provide systems, methods, and kits for CRISPR-based editing of DNA targets by a CRISPR-associated (Cas) enzyme. The systems include a bacterial host cell adapted to produce an engineered bacteriophage comprising a Cas protein and guide RNA that do not naturally occur together, i.e. they are engineered to occur together, as well as a DNA repair template comprising a donor DNA having a desired mutation. The guide RNA comprises a trans-activating crRNA and a guide sequence complementary to a target protospacer in a bacteriophage genome. A wild-type bacteriophage or a glucosylhydroxymethyl cytosine (ghmC)-unmodified mutant bacteriophage may be delivered into a disclosed bacterial host cell to create recombinants of bacteriophage having the desired mutation provided by the donor DNA.

Abstract: Techniques from two basic approaches, structure-based immunogen design and phage T4 nanoparticle delivery, are developed to construct new plague vaccines. The NH2-terminal ?-strand of F1 of Yersinia pestis is transplanted to the COOH-terminus of F1 of Yersinia pestis and the NH2-terminus sequence flanking the ?-strand of F1 of Yersinia pestis is duplicated to eliminate polymerization but to retain the T cell epitopes. The mutated F1 is fused to the V antigen of Yersinia pestis to thereby form a fusion protein F1mut-V mutant, which produces a completely soluble monomer. The fusion protein F1mut-V is then arrayed on phage T4 nanoparticles via a small outer capsid protein, Soc, from a T4 phage or a T4-related phage. Both the soluble and T4 decorated F1mut-V provided approximately 100% protection to mice and rats against pneumonic plague evoked by high doses of Yersinia pestis CO92.

Abstract: Techniques from two basic approaches, structure-based immunogen design and phage T4 nanoparticle delivery, are developed to construct new plague vaccines. The NH2-terminal ?-strand of F1 of Yersinia pestis is transplanted to the COOH-terminus of F1 of Yersinia pestis and the NH2-terminus sequence flanking the ?-strand of F1 of Yersinia pestis is duplicated to eliminate polymerization but to retain the T cell epitopes. The mutated F1 is fused to the V antigen of Yersinia pestis to thereby form a fusion protein F1mut-V mutant, which produces a completely soluble monomer. The fusion protein F1mut-V is then arrayed on phage T4 nanoparticles via a small outer capsid protein, Soc, from a T4 phage or a T4-related phage. Both the soluble and T4 decorated F1mut-V provided approximately 100% protection to mice and rats against pneumonic plague evoked by high doses of Yersinia pestis CO92.