Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells
Hannah I. Ghasemi, Julien Bacal, Amanda C. Yoon, Katherine U. Tavasoli, Carmen Cruz, Jonathan T. Vu, Brooke M. Gardner, Chris D. Richardson
Abstract
We describe a strategy to boost the efficiency of gene editing via homology-directed repair (HDR) by covalently modifying the template DNA with interstrand crosslinks. Crosslinked templates (xHDRTs) increase Cas9-mediated editing efficiencies by up to fivefold in K562, HEK293T, U2OS, iPS and primary T cells. Increased editing from xHDRTs is driven by events on the template molecule and requires ataxia telangiectasia and Rad3-related (ATR) kinase and components of the Fanconi anemia pathway.
Topics & Concepts
Genome editingDNAHomology directed repairFanconi anemiaDNA repairGeneTemplateBiologyChemistryGeneticsCRISPRMolecular biologyCell biologyNucleotide excision repairMaterials scienceNanotechnologyCRISPR and Genetic EngineeringDNA Repair MechanismsVirus-based gene therapy research