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AssigneeCaribou Biosciences, Inc.
InventorsPaul Daniel Donohoue, Andrew Paul May
Estimating the term…
The present specification discloses engineered Type II CRISPR-Cas9 systems comprising split-nexus Cas9-associated polynucleotides (sn-casPNs), including systems comprising three split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN/sn3-casPN) and systems comprising two split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN). Together with a Cas9 protein, the sn-casPNs facilitate site-specific modifications, including cleavage and mutagenesis, of a target polynucleotide in vitro and in vivo. Furthermore, the engineered Type II CRISPR-Cas9 systems comprising sn-casPNs are useful in methods of regulating expression of a target nucleic acid. Methods are described herein for the creation of a variety of engineered Type II CRISPR-Cas9 systems comprising two or more sn-casPNs. Polynucleotide sequences, expression cassettes, vectors, compositions, and kits for carrying out a variety of methods are also described. Furthermore, the present specification provides genetically modified cells, compositions of modified cells, transgenic organisms, pharmaceutical compositions, as well as a variety of compositions and methods involving the engineered Type II CRISPR-Cas9 systems.
One or more expression cassettes comprising:
The one or more expression cassettes of 13, wherein the one or more regulatory sequences facilitate expression of the Type II CRISPR-Cas9-associated polynucleotide composition and the Cas9 protein in a eukaryotic cell.
The one or more expression cassettes of 1, wherein the one or more regulatory sequences facilitate expression of the Type II CRISPR-Cas9-associated polynucleotide composition in a eukaryotic cell.
Dependencies are inferred from the claim text.