Integration and Stable Germ Line Transmission of Genes Injected into Mouse Pronuclei

Abstract

Genetic material has been successfully transferred into the genomes of newborn mice by injection of that material into pronuclei of fertilized eggs. Initial results indicated two patterns of processing the injected DNA: one in which the material was not integrated into the host genome, and another in which the injected genes became associated with high molecular weight DNA. These patterns are maintained through further development to adulthood. The evidence presented indicates the covalent association of injected DNA with host sequences, and transmission of such linked sequences in a Mendelian distribution to two succeeding generations of progeny.

Efficient Enhancement of Lentiviral Transduction Efficiency in Murine Spermatogonial Stem Cells

“…These techniques have allowed creation of transgenic animals via genetic modification and transplantation of SSCs that can pass on their traits to the progeny. The use of SSCs is more advantageous compared with pronuclear embryos or embryonic stem cells (ES cell) where the germ-line transmission is only 40-50% (Gordon and Ruddle, 1981). Therefore, the transgenic animal production using SSCs and the transplantation technique can be used to enhance the production efficiency of transgenic animals.…”

Evaluation of laser-assisted lentiviral transgenesis in bovine

“…The term "transgene" was first used for animals, which had integrated in vitro recombinated gene constructs into their genome (Gordon and Ruddle, 1981).…”

“…Injection of DNA into pronuclei of zygotes has been consistently used in mice since 1981 (Gordon and Ruddle, 1981) and in farm animals since 1985, when Hammer et al (1985) and Brem et al (1985) demonstrated the feasibility of this procedure.…”

Insights on bovine genetic engineering and cloning

“…Over the last decades several techniques were developed in order to produce genetically modified animals, and three of them became more popular due to their simplicity and positive results: exogenous pronuclear DNA microinjection in zygotes, injection of genetically modified embryonic stem (ES) cells into blastocysts, and, more recently, retrovirus mediated gene transfer (Gordon & Ruddle 1981, Hammer et al 1985, Gossler et al 1986, Hofmann et al 2003, 2004. Injection of ES cells into blastocysts is the most popular methodology of genetic manipulation in mice, once homologous recombination of ES cells in mice is feasible, allowing the contribution of modified cells in the embryos and therefore obtaining transgenic animals after breeding (Misra & Duncan 2002 Transgenic technology has become an essential tool for the development of animal biotechnologies, and animal cloning through somatic cell nuclear transfer (SCNT) enabled the generation of genetically modified animals utilizing previously modified and selected cell lineages as nuclei donors, assuring therefore the generation of homogeneous herds expressing the desired modification.…”