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PMID: 24918038 Published · epublish English Journal Article

Lentiviral vector transduction of spermatozoa as a tool for the study of early development.

FEBS open bio ·Vol. 4 ·2014-00-00 ·Pages 266-75

Chandrashekran A, Isa I, Dudhia J, Thrasher AJ, Dibb N, Casimir C, Readhead C, Winston R

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.

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
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Chandrashekran Anil
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.
Isa Ihsan
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.
Dudhia Jayesh
Department of Clinical Science Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts AL9 7TA, UK.
Thrasher Adrian J
Molecular Immunology Unit, UCL-Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK.
Dibb Nicholas
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.
Casimir Colin
Department of Natural Sciences, School of Science & Technology, Middlesex University, The Burroughs, London NW4 4BT, UK.
Readhead Carol
Biological Imaging Center, California Institute of Technology, Beckman Institute, 139-74, Pasadena, CA 91135, USA.
Winston Robert
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.
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Article Info
Journal
FEBS open bio
Abbr.
FEBS Open Bio
ISSN
2211-5463
DOI
10.1016/j.fob.2014.02.008
Published
2014-00-00
Epub
2014-00-03
Pages
266-75
Language
English
Region
England
NLM ID
101580716
PMCID
PMC4048842
External Links
PubMed source DOI: 10.1016/j.fob.2014.02.008 PMC Full Text View journal details
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