Crispr enzymes and systems
AssigneesThe Broad Institute, Inc., Massachusetts Institute of Technology
InventorsFeng Zhang, Aaron Smargon, Neena Pyzocha, David Benjamin Turitz Cox, Eric S. Lander
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Abstract
The invention provides for systems, methods, and compositions for targeting nucleic acids. In particular, the invention provides non-naturally occurring or engineered DNA or RNA-targeting systems comprising a novel DNA or RNA-targeting CRISPR effector protein and at least one targeting nucleic acid component like a guide RNA.
Claims26
A non-naturally occurring or engineered composition consisting of
- i) a Type VI-B CRISPR-Cas effector protein, and
- ii) a Type VI-B CRISPR-Cas recombinant guide RNA,
- wherein the recombinant guide RNA comprises, from 5′ to 3′ orientation, a) a heterologous guide sequence that is capable of hybridizing to a target RNA sequence, and b) a direct repeat sequence,
- whereby there is formed a CRISPR complex comprising the Type VI-B CRISPR-Cas effector protein complexed with the guide sequence that is hybridized to the target RNA sequence, and wherein the Type VI-B CRISPR-Cas effector protein and Type VI-B CRISPR-Cas guide RNA do not naturally occur together.
A non-naturally occurring or engineered composition consisting of
- i) a Type VI-B CRISPR-Cas effector protein,
- ii) a Type VI-B CRISPR-Cas recombinant guide RNA, and
- iii) a Type VI-B CRISPR-Cas accessory protein that enhances Type VI-B CRISPR-Cas effector protein activity, or a Type VI-B CRISPR-Cas accessory protein that represses Type VI-B CRISPR-Cas effector protein activity,
- wherein the recombinant guide RNA comprises, from 5′ to 3′ orientation, a) a heterologous guide sequence that is capable of hybridizing to a target RNA sequence, and b) a direct repeat sequence, whereby there is formed a CRISPR complex comprising the Type VI-B CRISPR-Cas effector protein complexed with the guide sequence that is hybridized to the target RNA sequence, and wherein the Type VI-B CRISPR-Cas effector protein and Type VI-B CRISPR-Cas guide RNA do not naturally occur together.
A non-naturally occurring or engineered composition consisting of
- i) a Type VI-B CRISPR-Cas effector protein having one or more nuclear localization signals and one or more functional domains covalently associated with the Type VI-B CRISPR-Cas effector protein, and
- ii) a Type VI-B CRISPR-Cas recombinant guide RNA,
- wherein the recombinant guide RNA comprises, from 5′ to 3′ orientation, a) a heterologous guide sequence that is capable of hybridizing to a target RNA sequence, and b) a direct repeat sequence, whereby there is formed a CRISPR complex comprising the Type VI-B CRISPR-Cas effector protein complexed with the guide sequence that is hybridized to the target RNA sequence, and wherein the Type VI-B CRISPR-Cas effector protein and Type VI-B CRISPR-Cas guide RNA do not naturally occur together.
Dependencies are inferred from the claim text.