title: High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases.
abstract: Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted genomic deletion of up to 100 kb and (iii) targeted insertion of small genetic elements concomitant with large genomic deletions.
authors.affiliation: Sigma-Aldrich Biotechnology, St. Louis, Missouri, USA.
authors.forename: Fuqiang
authors.initials: F
authors.lastname: Chen
authors.forename: Shondra M
authors.initials: SM
authors.lastname: Pruett-Miller
authors.forename: Yuping
authors.initials: Y
authors.lastname: Huang
authors.forename: Monika
authors.initials: M
authors.lastname: Gjoka
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Duda
authors.forename: Jack
authors.initials: J
authors.lastname: Taunton
authors.forename: Trevor N
authors.initials: TN
authors.lastname: Collingwood
authors.forename: Morten
authors.initials: M
authors.lastname: Frodin
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Davis
mesh_terms: D002460:Cell Line; D004277:DNA, Single-Stranded / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D016254:Mutagenesis, Insertional / Q000379:methods; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics
chemical_list: D004277:DNA, Single-Stranded; D009838:Oligodeoxyribonucleotides; D004720:Endonucleases