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US 12,606,834 B2Granted
Methods and compositions for efficient delivery of nucleic acids and rna-based antimicrobials
AssigneeNorth Carolina State University
InventorsChase Lawrence Beisel, Ahmed Abdelshafy Mahmoud Gomaa, Michelle Luo
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Abstract
The invention relates to the methods for modifying the methylation pattern of bacteriophage DNA and phagemid DNA and to methods for selective killing of bacteria using lysogenic bacteriophages comprising bacteriophage DNA or phagemid DNA comprising components of an engineered CRISPR-Cas system.
Claims13
A recombinant bacteriophage comprising:
- (a) a Type I CRISPR array comprising:
- i. a spacer sequence substantially complementary to a target DNA sequence of a target bacterium, wherein the target DNA sequence is adjacent to a protospacer adjacent motif (PAM), and
- ii. at least two repeat sequences;
- and
- (b) an exogenous nucleic acid construct encoding an exogenous Cas3 polypeptide that recognizes a complex of Cascade and the Type I CRISPR array, and an exogenous nucleic acid construct encoding a Cascade polypeptide that recognizes the Type I CRISPR array,
- (c) wherein the Type I CRISPR array, the exogenous nucleic acid construct encoding an exogenous Cas3 polypeptide, and/or the exogenous nucleic acid construct encoding a Cascade polypeptide is integrated into the bacteriophage DNA at a dispensable site or a complemented site.
3 more elements
Dependencies are inferred from the claim text.