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US 12,606,823 B2Granted
Modified CPF1 guide RNA
AssigneeBreeze Bio, Inc.
InventorKunwoo Lee
Life of this patent
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Abstract
The invention provides a nucleic acid comprising a Cpf1 crRNA, a processing sequence 5′ of the Cpf1 crRNA, and an extension sequence 5′ of the processing sequence. The invention also provides a composition comprising the nucleic acid, a carrier, and optionally Cpf1. Additionally, the invention provides method of genetically modifying a eukaryotic target cell, comprising contacting the eukaryotic target cell with the nucleic acid or the composition to genetically modify a target nucleic acid in the cell.
Claims29
A nucleic acid comprising
- a Cpf1 crRNA, wherein the Cpf1 crRNA comprises a stem-loop domain located 5′ of a targeting sequence;
- an extension sequence of at least 6 nucleotides 5′ of the crRNA;
- and, optionally, a processing sequence between the crRNA and the extension sequence, wherein the processing sequence is a sequence that is self-cleaved by Cpf1;
- wherein the extension sequence does not comprise a processing sequence, an aptamer, or the sequence of the Cpf1 crRNA, and wherein the extension sequence comprises a nucleotide comprising a 2′ deoxy modification.
1 more element
A composition comprising
- (a) a nucleic acid comprising a Cpf1 crRNA, wherein the Cpf1 crRNA comprises a stem-loop domain located 5′ of a targeting sequence;
- an extension sequence of at least 4 nucleotides and no more than 100 nucleotides positioned 5′ of the crRNA;
- and, optionally, a processing sequence between the crRNA and the extension sequence, wherein the processing sequence is a sequence that is self-cleaved by Cpf1;
- wherein the extension sequence does not comprise a processing sequence, an aptamer, or the sequence of the Cpf1 crRNA, and wherein the extension sequence comprises a nucleotide comprising a 2′ deoxy modification;
- and
- (b) a liposome, polymer nanoparticle, or metal nanoparticle;
- wherein the nucleic acid is in the liposome, or partially or totally encapsulated by the polymer nanoparticle, or attached to the metal nanoparticle or polymer nanoparticle.
4 more elements
A composition comprising
- (a) a nucleic acid comprising a Cpf1 crRNA, wherein the Cpf1 crRNA comprises a stem-loop domain located 5′ of a targeting sequence;
- an extension sequence of at least 4 nucleotides and no more than 100 nucleotides positioned 5′ of the crRNA;
- and, optionally, a processing sequence between the crRNA and the extension sequence, wherein the processing sequence is a sequence that is self-cleaved by Cpf1;
- wherein the extension sequence does not comprise a processing sequence, an aptamer, or the sequence of the Cpf1 crRNA;
- and
- (b) a Cpf1 protein;
- the nucleic acid and Cpf1 protein are in a liposome or partially or totally encapsulated by a polymer nanoparticle, or attached to a metal or polymer nanoparticle.
4 more elements
Dependencies are inferred from the claim text.