engleski

Gene trap mutagenesis of three genes expressed during mouse embryo development

After recent completion of the sequence of the human genome a wide quest for gene function is one of the current scientific priorities. One approach to assess the gene function in vivo is to create a mouse deficient for the gene of interest. In order to introduce genetic modification within the genome, and in the same step to produce the mutant mouse, the gene trap method was applied. The method comprises of random integration of a DNA vector containing promoterless lacZ and neoR fused genes. After selection of ES cells, mutant animals were produced and genes of interest selected according to the expression pattern in the embryos. Three mouse lines were selected in order to find genes important for embryo development. The genes were identified by rapid amplification of cDNA ends (RACE) in both 5’ and 3’ direction. Two of the genes were already known genes Nol1 and STAM2. The obtained sequence of the third gene was novel, and the gene was named kosenic. The active transcripts of the promoterless genes were shown to be located within endogenous genes expressed during embryo development. Three mutated mouse lines with integration in the three genes represent a tool to analyze expression and function of the modified genes.

mouse; embryo; gene trap

nije evidentirano