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US 2019/0100775 A1Published application
Engineered nucleic acid-targeting nucleic acids
AssigneeCaribou Biosciences, Inc.
InventorsPaul Daniel Donohoue, Andrew Paul May
Life of this patent
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Abstract
The present disclosure provides engineered polynucleotide sequences that form scaffolds and nucleoprotein complexes comprising such engineered polynucleotide sequences that form scaffolds and nucleic acid binding proteins. Nucleic acid sequences encoding the engineered polynucleotide sequences that form scaffolds, as well as expression cassettes, vectors and cells comprising such polynucleotide sequences, are described. A variety of methods for making and using the engineered polynucleotide sequences that form scaffolds are also disclosed.
Claims21
1-20. (canceled)
A cell comprising:
- a composition of engineered Class 2 Type II CRISPR-nucleic acid sequences forming a scaffold (“NASC”), the Class 2 Type II CRISPR-NASC composition comprising,
- a first engineered nucleic acid component (“NASC-PC1”) comprising, in a 5′ to 3′ direction,
- a spacer element 1 comprising a nucleic acid target binding sequence 1,
- a repeat element 1 comprising a repeat nucleic acid sequence 1, and
- a nucleic acid binding protein binding element 1 comprising a double-stranded nucleic acid binding protein binding sequence 1,
- wherein the spacer element 1 is covalently connected with the repeat element 1, and the repeat element 1 is covalently connected with the nucleic acid binding protein binding element 1;
- and
- a second engineered nucleic acid component (“NASC-PC2”) comprising, in a 5′ to 3′ direction,
- a spacer element 2 comprising a nucleic acid target binding sequence 2,
- a repeat element 2 comprising a repeat nucleic acid sequence 2, and
- a nucleic acid binding protein binding element 2 comprising a double-stranded nucleic acid binding protein binding sequence 2,
- wherein the spacer element 2 is covalently connected with the repeat element 2, and the repeat element 2 is covalently connected with the nucleic acid binding protein binding element 2;
- wherein there is a connection between the repeat nucleic acid sequence 1 and the repeat nucleic acid sequence 2 through hydrogen-bonded base pairs, the connection forms the Class 2 Type II CRISPR-NASC composition, and the Class 2 Type II CRISPR-NASC composition is capable of binding a first Class 2 Type II CRISPR-Cas9 protein and a second Class 2 Type II CRISPR-Cas9 protein.
10 more elements
Dependencies are inferred from the claim text.