High efficiency DNA delivery into swine oocytes and embryos by electronic pulse delivery (EPD).
The production of transgenic swine for xenotransplantation has been proposed as an optimal option to overcome the chronic shortage of human organ donors. Generation of genetically engineered swine has been elusive due to the difficulties in gene transfer. In order to achieve effective gene delivery, a key step for the genetic modification, we applied electronic pulse delivery (EPD) technology to introduce H2Kb-DC DNA construct into swine eggs. Using the developed EPD Protocols, we have achieved good viability of the EPD treated oocytes, satisfactory embryonic development of the EPD treated embryos, and stable DNA transfer into the swine embryos with high efficiency. Thus, application of the EPD technology promises to effectively facilitate the generation of large transgenic mammals.
Yang
Y
Y
InCell, Inc., Santa Clara, California 95051, USA. yyang@leland.stanford.edu
Huang
S H
SH
Zhao
X
X
D000818:Animals; D030801:Animals, Genetically Modified; D018957:CD59 Antigens / Q000235:genetics; D004247:DNA / Q000235:genetics; D004622:Embryo, Mammalian; D005314:Embryonic and Fetal Development; D018014:Gene Transfer Techniques; D006183:H-2 Antigens / Q000235:genetics; D006801:Humans; D009865:Oocytes; D011401:Promoter Regions, Genetic / Q000235:genetics; D013552:Swine / Q000235:genetics*; D019076:Transgenes / Q000235:genetics
D018957:CD59 Antigens; D006183:H-2 Antigens; C024755:H-2K(K) antigen; D004247:DNA