Double-strand break repair of eukaryotic DNA is mainly accomplished by non-homologous end joining and homologous recombination. Providing exogenous templates during homologous recombination repair can result in the editing of target genes, which is the central mechanism of the well-established CRISPR gene editing system. Currently, exogenous templates are mainly DNA molecules, which can provoke a cellular immune response within the cell. In order to verify the feasibility of RNA molecules as repair templates for homologous recombination in mammalian cell genome editing, we fused RNA template molecules to the 3´-end of sgRNA, so that the sgRNA and the homologous template RNA form a single RNA molecule. The results show this construct can be used as a repair template to achieve target gene editing in mammalian cells. In addition, the factors influencing homologous recombination mediated by RNA template molecules were investigated and it was found that increasing the length of homologous arms and inducing an R-loop near the DSB can effectively promote homologous recombination repair. Further, intracellular homologous chromosomes may compete with exogenous RNA templates. The findings in this paper provide a reference for the utilization of RNA template molecules to mediate target gene editing in eukaryotic cells, as well as a basis for the study of the mechanism by which RNA molecules mediate the repair of DSBs.