The aberrant expression level of intracellular
microRNAs (
miRNAs) holds great promise for differentiating cell types at the molecular level. However, cell subtype discrimination based on a single
miRNA molecular level is not sufficient and reliable. Herein, multiple identifiable and reporting modules are integrated into a single
DNA circuit for multiple
cancer cell subtypes identification based on
miRNAs bispecific recognition. The
DNA three-dimensional (3D) logic gate nano-hexahedron framework extends three recognition modules and three reporting modules to form three "AND" logic gates. Each Boolean operator "AND" returns an "ON" signal in the presence of bispecific
miRNAs, simultaneously enabling three types of cell subtype identification. It not only enables the discrimination of
cancer cells A549 and MCF-7 from normal cells NHDF but also successfully distinguishes different types of
cancer cells. The bispecific intracellular
miRNA controllable
DNA circuit, with low signal-to-noise ratio, easily extends to various cell type discrimination by adjusting the
miRNA species, provides huge opportunities for accurately differentiating multiple cell types at the molecular level.