Abilayer MOF reporter (ZIF-67@FAM-mRNA@ZIF-8) was synthesized, and the ZIF-67 was used as a carrier and fluorescent quencher to connect the FAM reporter through electrostatic adsorption and coordination effect. The ZIF-8 covering the outer layer can improve the stability of the probe and cell permeability, which helps the FAM reporter effectively release. After entering the cancer cells, the acidic environment in the cells induced the decomposition of ZIF-8. The excess ATP in the tumor cells competitively binds ZIF-67, causing the FAM reporter to shed and restore fluorescence. The shed FAM reporter was specifically bound to the overexpressed miRNA-21 in breast cancer cells to achieve fluorescence imaging and therapy of breast cancer. The results of specific imaging and apoptosis experiments of breast cancer cells indicate that bilayer MOF nanomachine provides an effective nanotherapy platform for accurate fluorescence imaging.