Inactivation of Apoe and Apoc1 by two consecutive rounds of gene targeting: effects on mRNA expression levels of gene cluster members.

The genes encoding apolipoprotein (apo) E and apoC1 are, together with the gene for apoC2, located in a conserved gene cluster on human chromosome 19q12-13.2 and mouse chromosome 7. Although the significance of apoE as a ligand for receptor-mediated uptake of lipoprotein remnant particles is undisputed, the in vivo function of apoC1 and the possible interaction between apoE and apoC1 in the modulation of plasma cholesterol and triglyceride levels is far from understood. Our strategy to unravel the metabolic relationship between apoE and apoC1 in vivo is to first generate mice deficient in both apolipoproteins, enabling future production of transgenic mice with variable ratios of normal and mutant apoE and apoC1 on a null background. Here we report the creation and characterization of mice deficient in both apoE and apoC1. As these genes are tightly genetically linked, double-deficient mice were obtained by two consecutive rounds of gene targeting in mouse embryonic stem cells. Surprisingly, double inactivation of the Apoe and Apoc1 gene loci as well as single inactivations at either one of these loci were found to affect also the RNA expression levels of the other gene members in the Apoe-c1-c2 cluster. This indicates that targeted insertions are not necessarily neutral for the expression of nearby gene members in a given gene cluster. Homozygous Apoe-c1 knockout mice are hypercholesterolemic, with serum cholesterol levels of 12.5 +/- 4.3 mM compared with 2.9 +/- 0.5 mM in control mice, resembling mice solely deficient in apoE.