Constructs for improved HDR-dependent genomic editing
AssigneesThe Broad Institute, Inc., President and Fellows of Harvard College
InventorsDavid R. Liu, Holly A. Rees
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Abstract
The invention provides an improved genome editing construct which is capable of editing a target sequence in an HDR-dependent manner (i.e., “HDR-dependent genome editors”) with increased efficiency and reduced indel formation and which does not require a dividing cell. In particular, the instant specification provides a new fusion protein comprising a nucleic acid programmable DNA binding protein (napDNAbp) (e.g., Cas9) with a nickase activity and a single-stranded DNA binding protein (e.g., Rad51) which edits a target DNA in an HDR-dependent manner with greater efficiency (e.g., increased rate of induced HDR) and/or with a lower rate or occurrence of indel formation.
Claims21
A fusion protein comprising (a) a nucleic acid programmable DNA binding protein (napDNAbp) comprising a nickase activity, and (b) a Rad51 variant comprising an S208E mutation and an A209D mutation relative to a wild type Rad51 protein, wherein the fusion protein is capable of inducing homology-directed repair with a lower rate of indel formation as compared to the rate of indel formation induced by a control protein comprising a napDNAbp comprising a nickase activity that is not fused to a Rad51 variant comprising an S208E mutation and an A209D mutation relative to a wild type Rad51 protein.
Dependencies are inferred from the claim text.