title: Lentiviral vector transduction of spermatozoa as a tool for the study of early development.
abstract: Spermatozoa and lentiviruses are two of nature's most efficient gene delivery vehicles. Both can be genetically modified and used independently for the generation of transgenic animals or gene transfer/therapy of inherited disorders. Here we show that mature spermatozoa can be directly transduced with various pseudotyped lentiviral vectors and used in in vitro fertilisation studies. Lentiviral vectors encoding Green Fluorescent Protein (GFP) were shown to be efficiently processed and expressed in sperm. When these transduced sperm were used in in vitro fertilisation studies, GFP expression was observed in arising blastocysts. This simple technique of directly transducing spermatozoa has potential to be a powerful tool for the study of early and pre-implantation development and could be used as a technique in transgenic development and vertical viral transmission studies.
authors.affiliation: Department of Surgery and Cancer, Division of Cancer, Imperial College London, Hammersmith Campus, Institute of Reproductive and Developmental Biology (IRDB), Du Cane Road, London W12 0NN, UK.
authors.forename: Anil
authors.initials: A
authors.lastname: Chandrashekran
authors.affiliation: Department of Surgery and Cancer, Division of Cancer, Imperial College London, Hammersmith Campus, Institute of Reproductive and Developmental Biology (IRDB), Du Cane Road, London W12 0NN, UK.
authors.forename: Ihsan
authors.initials: I
authors.lastname: Isa
authors.affiliation: Department of Clinical Science Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts AL9 7TA, UK.
authors.forename: Jayesh
authors.initials: J
authors.lastname: Dudhia
authors.affiliation: Molecular Immunology Unit, UCL-Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.affiliation: Department of Surgery and Cancer, Division of Cancer, Imperial College London, Hammersmith Campus, Institute of Reproductive and Developmental Biology (IRDB), Du Cane Road, London W12 0NN, UK.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Dibb
authors.affiliation: Department of Natural Sciences, School of Science & Technology, Middlesex University, The Burroughs, London NW4 4BT, UK.
authors.forename: Colin
authors.initials: C
authors.lastname: Casimir
authors.affiliation: Biological Imaging Center, California Institute of Technology, Beckman Institute, 139-74, Pasadena, CA 91135, USA.
authors.forename: Carol
authors.initials: C
authors.lastname: Readhead
authors.affiliation: Department of Surgery and Cancer, Division of Cancer, Imperial College London, Hammersmith Campus, Institute of Reproductive and Developmental Biology (IRDB), Du Cane Road, London W12 0NN, UK.
authors.forename: Robert
authors.initials: R
authors.lastname: Winston
keywords: 293T, Human embryonic kidney cells; 7-AAD, 7-Aminoactinomycin D; AZT, azidodeoxythimidine; CMV, Cytomegalovirus promoter; Development; EF-1, Elongation factor 1 alpha promoter; GFP, Green Fluorescent Protein; IVF, in vitro fertilisation; In vitro fertilisation; LTR, Long Terminal Repeat; Lentiviral vectors; PGK, Phosphoglycerate kinase promoter; Spermatozoa; Transduction; Transgenics; UCOE, ubiquitous chromatin opening element promoter; VSV-g, vesicular stomatitis virus