Lentiviral vector transduction of spermatozoa as a tool for the study of early development
Anil Chandrashekran,
Ihsan Isa,
Jayesh Dudhia,
Adrian J. Thrasher,
Nicholas Dibb,
Colin Casimir,
Carol Readhead,
Robert Winston
Affiliations
Anil Chandrashekran
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
Ihsan Isa
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
Jayesh Dudhia
Department of Clinical Science Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts AL9 7TA, UK
Adrian J. Thrasher
Molecular Immunology Unit, UCL-Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK
Nicholas Dibb
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
Colin Casimir
Department of Natural Sciences, School of Science & Technology, Middlesex University, The Burroughs, London NW4 4BT, UK
Carol Readhead
Biological Imaging Center, California Institute of Technology, Beckman Institute, 139-74, Pasadena, CA 91135, USA
Robert Winston
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
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.
