Compositions and methods of nucleic acid-targeting nucleic acids
AssigneeCaribou Biosciences, Inc.
InventorsAndrew Paul May, Rachel E. Haurwitz, Jennifer A. Doudna, James M. Berger, Matthew Merrill Carter, Paul Daniel Donohoue
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Abstract
This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.
Claims15
A method of delivering a donor polynucleotide to a target nucleic acid within a nucleus of a cell, the method comprising contacting the cell with a complex comprising:
- a. a site-directed polypeptide fused to a reverse transcriptase, and
- b. an engineered single guide nucleic acid-targeting nucleic acid (NATNA), the engineered single guide NATNA comprising a spacer that can specifically hybridize to the target nucleic acid, a crRNA, a tracrRNA, and a 3′ hybridizing extension comprising a reverse transcription template and a binding site for the reverse transcriptase, wherein said 3′ hybridizing extension is RNA,
- wherein the site-directed polypeptide introduces a single-stranded break within the target nucleic acid, and wherein the reverse transcription template is reverse transcribed by the reverse transcriptase within the nucleus to deliver a newly transcribed DNA comprising the donor polynucleotide to the nucleus of the cell.
Dependencies are inferred from the claim text.