Genetic transformation of mouse embryos by microinjection of purified DNA.

Abstract

A recombinant plasmid composed of segments of herpes simplex virus and simian virus 40 viral DNA inserted into the bacterial plasmid pBR322 was microinjected into pronuclei of fertilized mouse oocytes. The embryos were implanted in the oviducts of pseudopregnant females and allowed to develop to term. DNA from newborn mice was evaluated by the Southern blotting technique for the presence of DNA homologous to the injected plasmid. Two of 78 mice in one series of injections showed clear homology, though the injected sequences had been rearranged. Band intensities from the two positive mice were consistent with the presence of donor DNA in most or all of the cells of the newborns. These results demonstrate that genes can be introduced into the mouse genome by direct insertion into the nuclei of early embryos. This technique affords the opportunity to study problems of gene regulation and cell differentiation in a mammalian system by application of recombinant DNA technology.

Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing

“…The first transgenic mouse was produced by Gordon in 1980 [32]. By constructing transgenes that contained GFP cDNA driven by specific promoter sequences the target proteins could now be identified by detecting the green fluorescence [33].…”

Lamr1 functional retroposon causes right ventricular dysplasia in mice

“…We constructed three kinds of targeting vectors under the following promoters (Lamr1-tp1, KSCX and α-MHC; Lamr1, α-MHC). We introduced these targeting vectors into blastocysts (derived from the C57Bl/6 Jcl mouse strain) by a standard pronuclear microinjection technique 34 .…”

Status of transgenic chicken models for developmental biology

“…The common method for introducing genes into mammals is to microinject DNA into the pronucleus of a newly fertilized egg (Gordon et al, 1980;Brinster et al, 1981). However, direct DNA injection of freshly laid chicken eggs is much more technically difficult compared with mammals because a fertile freshly laid chicken egg contains approximately 50,000 -60,000 cells when it is laid (Spratt and Haas, 1960).…”