title: Sustained in vivo inhibition of protein domains using single-chain Fv recombinant antibodies and its application to dissect RGMa activity on axonal outgrowth.
abstract: Antibodies are powerful tools for delineating the specific function of protein domains, yet several limitations restrict their in vivo applicability. Here we present a new method to obtain sustained in vivo inhibition of specific protein domains using recombinant antibodies. We show that long term in vivo expression of single-chain Fv (scFv) fragments in the developing CNS can be achieved through retroviral transduction. Moreover, specific scFvs generated against the N- and C-terminal domains of the repulsive guidance molecule, RGMa, prevent proper axon targeting in the visual system. This work reveals a previously unappreciated role for the RGMa N-terminal domain in axon guidance, and provides a novel, broadly applicable and rapid procedure to functionally antagonize any protein domain in vivo.
authors.affiliation: Genetics and Development Division, Toronto Western Research Institute, Toronto, Ontario, Canada M5T 2S8.
authors.forename: Nardos G
authors.initials: NG
authors.lastname: Tassew
authors.forename: Jason
authors.initials: J
authors.lastname: Charish
authors.forename: Larisa
authors.initials: L
authors.lastname: Chestopalova
authors.forename: Philippe P
authors.initials: PP
authors.lastname: Monnier
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000494:pharmacology; D001369:Axons / Q000187:drug effects*; D002490:Central Nervous System / Q000166:cytology* / Q000196:embryology; D002642:Chick Embryo; D036386:Ephrin-A5 / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D000906:Antibodies; D036386:Ephrin-A5; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins