title: Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus.
abstract: We demonstrate enhanced transgenesis in mice by intracytoplasmic injection of envelope-free lentivirus. Envelope-free lentivirus carrying the green fluorescent protein (GFP) gene under the control of the ubiquitin promoter (LVU-GFP) was microinjected into the cytoplasm of mouse zygotes prior to embryo transfer. Ninety-seven percent (31/32) of the adult mice were confirmed transgenic by PCR and Southern blot analysis; all founder mice express GFP when tail snips were examined by fluorescent microscopy prior to genomic DNA extraction. Transgene insertion numbers ranging from 1 to 32 were revealed by Southern blot analysis. Germline transmission was confirmed by the presence of transgene in F1 offspring. As expected, a lower transgenic rate (2.2%; 1/46) resulted when envelope-free LVU-GFP was microinjected into the perivitelline space (PVS) because cell recognition followed by membrane fusion between the viral envelope and the target cell is prerequisite for successful infection by envelope viruses. Here we demonstrate the competence of envelope-free lentivirus in establishing stable gene integration by germline transgenesis in mice at high efficiency, by intracytoplasmic viral injection (INVI) of envelope-free lentivirus into mouse zygotes.
authors.affiliation: Yerkes National Primate Research Center, Emory University, Atlanta, Georgia 30329, USA.
authors.forename: Shang-Hsun
authors.initials: SH
authors.lastname: Yang
authors.forename: Yuksel
authors.initials: Y
authors.lastname: Agca
authors.forename: Pei-Hsun
authors.initials: PH
authors.lastname: Cheng
authors.forename: Jin-Jing
authors.initials: JJ
authors.lastname: Yang
authors.forename: Cansu
authors.initials: C
authors.lastname: Agca
authors.forename: Anthony Wing Sang
authors.initials: AW
authors.lastname: Chan
mesh_terms: D000818:Animals; D003593:Cytoplasm / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000235:genetics; D007267:Injections; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D015053:Zygote / Q000648:ultrastructure / Q000821:virology
chemical_list: D049452:Green Fluorescent Proteins