PRODUCTION OF FERTILE TRANSGENIC ZEBRAFISH FROM LONG-TERM CULTURED CELLS BY NUCLEAR TRANSFER

Shuo Lin
Department of Molecular,
Cell & Developmental Biology,
University of California, Los Angeles,
621 Charles E. Young Drive South, LS4325,
Los Angeles, CA 90095-1606
telephone: 310-267-4970
fax: 310-267-4971
email: shuolin@ucla.edu
prestype: Platform
presenter: Shuo Lin

Ki-Young Lee #*, Haigen Huang #, Jae Yong Han*, and Shuo Lin #

# Department of Molecular, Cell & Developmental Biology, University of California, Los Angeles, 621 Charles E. Young Drive South, LS4325, Los Angeles, CA 90095-1606
* School of Agriculture Biotechnology, Seoul National University, Korea

ABSTRACT

The zebrafish has become an important model for genetic and developmental biology studies. However, due to the lack of embryonic stem cells, one major limitation of this model is the inability to perform site-specific genetic modifications. In some other animals, generation of gene knockout by somatic cell cloning has been used to overcome this limitation. As the first step towards this goal, we established conditions to produce cloned transgenic zebrafish using a long-term cultured embryonic fibroblast cells. First, these cells were infected with a retrovial vector expressing the GFP marker gene. The nuclei of these cells were then transplanted into enucleated, unfertilized eggs and the resultant cloned zebrafish were shown to be fertile and continue to express the GFP reporter gene. From 10 experimental groups, a total of 34 embryos (36.2 %) reached the blastula stage while 16 embryos (17.0%) from 6 groups hatched, and 11 individuals reached adulthood. All of these individuals produced offspring after mating with wild type, and their F1 and F2 progeny expressed GFP with similar expression patterns as the F0 generation. This study sets up a foundation for further genetic manipulations in zebrafish.