Construction of Rtn4-A/Bknockout mouse model

Objective To generate Rtn4-A/Bknockout mouse model and to explore the biological function of the Rtn4-B gene.Methods The targeting construct for inactivating Rtn4-A/Bgene was prepared by bacterial artificial chromosome(BAC).The vector was linearized and electroporated into 129SvEv mouse embryonic stem cells(ES cells).Then the Rtn4-A/Bknockout ES cells were microinjected into blastula of C57BL/6Jmice after superovulation.F1hybrid mice were bred to obtain mouse aggregation chimeras,and were identified by PCR amplification of tail genomic DNA.Results Fourteen clones of gene-targeted ES cells were identified after gene knockout and five male chimeras with a higher than 50chimeric ratio were produced after microinjection into the blastula.Finally four Rtn4-A/Bhybrid mice were obtained.Conclusion ARtn4-A/Bdeficient mouse strain has been successfully generated by homologous recombination using genetically modified ES cells.