Liposome-mediated DNA transfer into chicken primordial germ cells in vivo.

Abstract

In embryogenesis, avian primordial germ cells (PGCs) circulate temporarily in the blood vessels at stages 10-15 (Hamburger and Hamilton, 1951), before reaching the gonads. In an attempt to transfer cloned genes into PGCs, liposome consisting of reporter plasmid DNA and N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammoniummethylsulf ate was injected into the marginal veins of embryos at stages 11-15. As reporter plasmids, pRSVZ and pAcZ harboring the Escherichia coli lacZ gene driven, respectively, by the Rous sarcoma virus (RSV) promoter and the chicken beta-actin gene promoter were used. First, 55 embryos were injected with liposome containing pRSVZ and stained for the bacterial beta-galactosidase activity 24 hr after injection. In all the embryos, cells positive for beta-galactosidase activity were observed among the blood cells, endothelial cells, and endocardium cells of the heart, suggesting that transfection took place within the circulatory system. Then, embryos were injected with liposome containing pRSVZ or pAcZ, and stained 2 or 3 d after injection. PGCs positive for beta-galactosidase activity were observed in the gonads in four out of 44 embryos injected with pRSVZ, and 29 out of 71 embryos injected with pAcZ, indicating that the plasmid DNA was transferred into PGCs developing normally. The average number of positive PGCs per embryo was 0.2 and 2.1, respectively, when pRSVZ and pAcZ were introduced. The difference in the number of positive PGCs detected after introduction of the two plasmids suggests that the actin promoter has a higher level of transcriptional activity in PGCs than does the RSV promoter.