Complexes for gene deletion and editing
AssigneeUniversity of Massachusetts
InventorsMichael P. CZECH, Yuefei SHEN
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Abstract
Complexes comprising a nucleic acid-guided endonuclease, a sequence-specific targeting nucleic acid and an amphipathic helical peptide are provided. Compositions and methods for delivery of complexes comprising a nucleic acid-guided endonuclease, a sequence-specific targeting nucleic acid and an amphipathic helical peptide to mammals for both research and therapeutic use are provided. Methods of treating or reducing one or more symptoms of type 2 diabetes, prediabetes and/or gestational diabetes are provided.
Claims68
A composition comprising:
- a nucleic acid-guided endonuclease,
- a sequence-specific targeting nucleic acid and
- an amphipathic helical peptide,
- wherein the nucleic acid-guided endonuclease, the sequence-specific targeting nucleic acid and the amphipathic helical peptide form a complex, and
- wherein the amphipathic helical peptide mediates delivery of the complex to a target cell, and wherein the nucleic acid-guided endonuclease mediates editing or deletion of a target gene in the target cell.
The composition of any one of the previous claims, wherein the complex further comprises an aptamer molecule with binding specificity for the target cell.
The composition of any one of the previous claims, wherein the complex is encapsulated in a glucan particle (GP).
A method of editing or deleting a target gene in a cell comprising contacting a cell with a complex comprising a nucleic acid-guided endonuclease, a sequence-specific targeting nucleic acid and an amphipathic helical peptide and allowing the complex to enter the cell and edit or delete the target gene.
A method of improving glucose tolerance in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a complex including a nucleic acid-guided endonuclease, a sequence-specific targeting nucleic acid and an amphipathic helical peptide, wherein the complex edits or deletes a target gene in the subject.
The method of claim31, wherein the target gene is RIP140.
A method of improving glucose tolerance in a subject, comprising:
- contacting an adipocyte cell or pre-adipocyte cell ex vivo with a complex including a Cas9, a RIP140 guide RNA and an amphipathic helical peptide for a sufficient amount of time to delete or inactivate RIP140 gene in the cell;
- and
- implanting into the subject the cell having the deleted or inactivated RIP140 gene.
A guide RNA of between 15 and 30 bases in length, that comprises a region which is at least 95% homologous to SEQ ID NO:
- 1.
A guide RNA comprising SEQ ID NO:
- 1.
A guide RNA of between 15 and 30 bases in length, that comprises a region which is at least 95% homologous to SEQ ID NO:
- 2.
A guide RNA comprising SEQ ID NO:
- 2.
A guide RNA of between 15 and 30 bases in length, that comprises a region which is at least 95% homologous to SEQ ID NO:
- 3.
A guide RNA comprising SEQ ID NO:
- 3.
A guide RNA of between 15 and 30 bases in length, that comprises a region which is at least 95% homologous to SEQ ID NO:
- 4.
A guide RNA comprising SEQ ID NO:
- 4.
Dependencies are inferred from the claim text.