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US 2023/0407295 A1Published application
Protecting oligonucleotides for crispr guide rna
AssigneeUniversity of Massachusetts
InventorsHan Zhang, Erik Joseph Sontheimer, Jonathan Kenneth Watts, Anastasia Khvorova
Life of this patent
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Abstract
Protecting oligonucleotides are provided. Protecting oligonucleotides with 5′ and/or 3′ conjugated moieties are provided. Protecting oligonucleotides with chemical modifications are provided. Methods of using the protecting oligonucleotides for genome editing with a CRISPR nuclease and kits for performing the same are also provided.
Claims28
A protecting oligonucleotide comprising:
- (a) a sequence that is complementary to a crRNA;
- and
- (b) at least one chemically modified nucleotide,
- wherein the protecting oligonucleotide is capable of binding the crRNA, and
- wherein the protecting oligonucleotide confers nuclease resistance to the crRNA when bound.
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A double stranded oligonucleotide comprising:
- (a) a crRNA comprising (i) a guide sequence portion capable of hybridizing to a target polynucleotide sequence, and (ii) a repeat sequence portion;
- and
- (b) a protecting oligonucleotide that is complementary to the crRNA,
- wherein the crRNA comprises at least 50% modified nucleotides, and
- wherein the protecting oligonucleotide comprises at least one modified nucleotide.
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Dependencies are inferred from the claim text.