Sperm-mediated gene transfer studies on zebrafish in Singapore

This paper describes the work conducted in the Department of Biological Sciences, National University of Singapore; on sperm-mediated gene transfer in zebrafish. This study began by carrying out direct interaction experiments of pUSVCAT reporter DNA with spermatozoa. Other constructs, including pXGH5, pMTL, pRSVL and pGEM-luc, were subsequently used. The different constructs were taken up by sperm cells with comparable efficiencies. In general, no reporter gene expression, Mendelian inheritance, or evidence of genomic integration of the foreign sequences were obtained. However, transmission of the reporter DNA through generations was observed. DNA uptake by sperm cells was shown using FISH and was enhanced by electroporation. The potential use of more recent approaches, such as REMI and ICSI are explored. Future research directions are discussed.