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

Creators: Chandrashekran, Anil; Isa, Ihsan; Dudhia, Jayesh; Thrasher, Adrian J.; Dibb, Nicholas; Casimir, Colin; Readhead, Carol; Winston, Robert

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.

Additional Information

© 2014 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/). Received 2 October 2013, Revised 13 February 2014, Accepted 19 February 2014, Available online 3 March 2014. We thank Ms. Rochelle Diamond (Caltech) for help with FACS analysis. Thanks to Dr. Michael Harkey, Fred Hutchinson Cancer Center, USA for LAM-PCR analysis. Thanks also due to Drs. Mark Fenwick and Jocelyn Mora, Imperial College London for help with confocal microscopy. This work was supported by Atazoa Ltd and, in part the Genesis Research Trust previously known as the Institute of Obstetrics and Gynaecology Trust, UK and Wellcome Trust, UK. Authorship contributions: AC, designed, performed experiments, analysed the data and wrote the paper; II, performed experiments; J.D., A.T., N.D., C.C., R.W., C.R., assisted in the design, performed experiments and contributed to the writing of the paper.

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