Cloned zebrafish by nuclear transfer from long-term-cultured cells.
Although mammals have been cloned from genetically manipulated cultured cells, a comparable achievement has not been realized in lower vertebrates. Here we report that fertile transgenic zebrafish can be obtained by nuclear transfer using embryonic fibroblast cells from long-term cultures. The donor nuclei, modified by retroviral insertions expressing green fluorescent protein (GFP), were transplanted into manually enucleated eggs. Overall, a 2% success rate was achieved, resulting in 11 adult transgenic zebrafish expressing GFP. These nuclear transplants produced fertile, diploid offspring, and their F1/F2 progeny continued to express GFP in a pattern identical to that of the founder fish. This finding demonstrates that slowly dividing nuclei from cultured cells can be reprogrammed to support rapid embryonic development and sets up a foundation for targeted genetic manipulation in zebrafish.
Lee
Ki-Young
KY
Department of Molecular, Cellular, and Developmental Biology, University of California Los Angeles, Los Angeles, CA 90095-1606, USA.
Huang
Haigen
H
Ju
Bensheng
B
Yang
Zhongan
Z
Lin
Shuo
S
D000818:Animals; D002478:Cells, Cultured; D002875:Chromosomes / Q000235:genetics; D019976:Cloning, Organism / Q000379:methods*; D004625:Embryo, Nonmammalian / Q000196:embryology*; D047108:Embryonic Development; D005260:Female; D005347:Fibroblasts; D008297:Male; D053652:Nuclear Transfer Techniques; D010063:Ovum / Q000166:cytology; D013997:Time Factors; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000254:growth & development