title: Coupling endonucleases with DNA end-processing enzymes to drive gene disruption.
abstract: Targeted DNA double-strand breaks introduced by rare-cleaving designer endonucleases can be harnessed for gene disruption applications by engaging mutagenic nonhomologous end-joining DNA repair pathways. However, endonuclease-mediated DNA breaks are often subject to precise repair, which limits the efficiency of targeted genome editing. To address this issue, we coupled designer endonucleases to DNA end-processing enzymes to drive mutagenic break resolution, achieving up to 25-fold enhancements in gene disruption rates.
authors.affiliation: Molecular and Cellular Biology Program, University of Washington, Seattle, Washington, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Certo
authors.forename: Kamila S
authors.initials: KS
authors.lastname: Gwiazda
authors.forename: Ryan
authors.initials: R
authors.lastname: Kuhar
authors.forename: Blythe
authors.initials: B
authors.lastname: Sather
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Curinga
authors.forename: Tyler
authors.initials: T
authors.lastname: Mandt
authors.forename: Michelle
authors.initials: M
authors.lastname: Brault
authors.forename: Abigail R
authors.initials: AR
authors.lastname: Lambert
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Baxter
authors.forename: Kyle
authors.initials: K
authors.lastname: Jacoby
authors.forename: Byoung Y
authors.initials: BY
authors.lastname: Ryu
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Kiem
authors.forename: Agnes
authors.initials: A
authors.lastname: Gouble
authors.forename: Frederic
authors.initials: F
authors.lastname: Paques
authors.forename: David J
authors.initials: DJ
authors.lastname: Rawlings
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Scharenberg
mesh_terms: D000818:Animals; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D004260:DNA Repair; D004720:Endonucleases / Q000502:physiology*; D005090:Exodeoxyribonucleases / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D010750:Phosphoproteins / Q000502:physiology; D019713:Receptors, CCR5 / Q000502:physiology
chemical_list: D010750:Phosphoproteins; D019713:Receptors, CCR5; D004720:Endonucleases; D005090:Exodeoxyribonucleases; C486104:TREX2 protein, human