title: Knockout rats via embryo microinjection of zinc-finger nucleases.
abstract: The toolbox of rat genetics currently lacks the ability to introduce site-directed, heritable mutations into the genome to create knockout animals. By using engineered zinc-finger nucleases (ZFNs) designed to target an integrated reporter and two endogenous rat genes, Immunoglobulin M (IgM) and Rab38, we demonstrate that a single injection of DNA or messenger RNA encoding ZFNs into the one-cell rat embryo leads to a high frequency of animals carrying 25 to 100% disruption at the target locus. These mutations are faithfully and efficiently transmitted through the germline. Our data demonstrate the feasibility of targeted gene disruption in multiple rat strains within 4 months time, paving the way to a humanized monoclonal antibody platform and additional human disease models.
authors.affiliation: Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI 52336, USA.
authors.forename: Aron M
authors.initials: AM
authors.lastname: Geurts
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Cost
authors.forename: Yevgeniy
authors.initials: Y
authors.lastname: Freyvert
authors.forename: Bryan
authors.initials: B
authors.lastname: Zeitler
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Vivian M
authors.initials: VM
authors.lastname: Choi
authors.forename: Shirin S
authors.initials: SS
authors.lastname: Jenkins
authors.forename: Adam
authors.initials: A
authors.lastname: Wood
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Cui
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Meng
authors.forename: Anna
authors.initials: A
authors.lastname: Vincent
authors.forename: Stephen
authors.initials: S
authors.lastname: Lam
authors.forename: Mieczyslaw
authors.initials: M
authors.lastname: Michalkiewicz
authors.forename: Rebecca
authors.initials: R
authors.lastname: Schilling
authors.forename: Jamie
authors.initials: J
authors.lastname: Foeckler
authors.forename: Shawn
authors.initials: S
authors.lastname: Kalloway
authors.forename: Hartmut
authors.initials: H
authors.lastname: Weiler
authors.forename: Séverine
authors.initials: S
authors.lastname: Ménoret
authors.forename: Ignacio
authors.initials: I
authors.lastname: Anegon
authors.forename: Gregory D
authors.initials: GD
authors.lastname: Davis
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Fyodor D
authors.initials: FD
authors.lastname: Urnov
authors.forename: Howard J
authors.initials: HJ
authors.lastname: Jacob
authors.forename: Roland
authors.initials: R
authors.lastname: Buelow
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA; D004622:Embryo, Mammalian; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D005240:Feasibility Studies; D005260:Female; D055786:Gene Knockout Techniques; D049452:Green Fluorescent Proteins; D007075:Immunoglobulin M / Q000235:genetics*; D008297:Male; D008845:Microinjections; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D012333:RNA, Messenger; D051381:Rats; D016335:Zinc Fingers / Q000235:genetics; D020691:rab GTP-Binding Proteins / Q000235:genetics*
chemical_list: D007075:Immunoglobulin M; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D004247:DNA; D004706:Endodeoxyribonucleases; C498787:Rab38 protein, rat; D020691:rab GTP-Binding Proteins