Components for genomic editing
AssigneeKano Therapeutics Inc.
InventorsGregory Benjamin Foley, Floris Alaska Silvia Engelhardt, Stephanie Barr
Life of this patent
Estimating the term…
Abstract
The present disclosure provides, among other things, homology directed repair (HDR) using single stranded DNA (ssDNA), e.g., circular single stranded DNA (cssDNA) and compositions for use in the same.
Claims60
A composition comprising a cssDNA molecule, wherein the cssDNA molecule comprises a co-location sequence, a selectable sequence, a packaging signal, and a designed sequence, wherein the co-location sequence is capable of associating with a synthetic nuclease.
A composition comprising:
- a cell, wherein the cell comprises
- a synthetic nuclease, wherein the synthetic nuclease is associated with a synthetic binding domain, wherein the synthetic binding domain is capable of associating with a co-location sequence, and
- a cssDNA molecule, wherein the cssDNA molecule comprises the co-location sequence.
A vector comprising a nucleic acid sequence that encodes a ZF5-Cas9 fusion protein comprising an amino acid sequence according to SEQ ID NO:
- 51 or 53.
A method of engineering a target dsDNA in a cell comprising:
- contacting a target dsDNA with a synthetic nuclease and a cssDNA comprising a co-location sequence;
- and culturing the cell.
A method of making a kit for editing dsDNA, comprising:
- selecting a cssDNA comprising a co-location sequence;
- selecting an synthetic nuclease wherein the synthetic nuclease comprises a synthetic binding domain and wherein the synthetic binding domain associates with the co-location sequence;
- and assembling the cssDNA and synthetic nuclease into a kit.
A fusion protein comprising a Cas9 nuclease linked to a zinc finger (ZF) via a flexible linker.
A complex comprising:
- a Cas nuclease linked to a ZF via a flexible linker or through a protein:protein interaction;
- and
- a cssDNA molecule comprising a designed sequence and a co-location sequence;
- wherein the co-location sequence comprises a ZF DNA binding motif (ZFDBM);
- and
- wherein the the ZF binds to the ZFDBM, thereby forming the complex.
Dependencies are inferred from the claim text.