This review discusses the construction, regulation, and application of chiral structures in azobenzene-containing systems. Chirality, a fundamental property in nature and biomolecules, plays a crucial role in various biological processes. The synthesis of chiral artificial structures is essential for understanding the origin of enantiomeric imbalance and its impact on biomolecular functions. Azobenzene (Azo) molecules, known for their reversible cis-trans isomerization, are widely used in functional dyes and materials due to their photoresponsive properties. The review highlights the dynamic regulation of chiroptical properties through temperature, solvent, magnetic field, and pH, enabling the construction of stimuli-responsive chiral nanomaterials with diverse functionalities. The construction of chiral structures in Azo-containing systems involves both chiral and achiral components. Chiral Azo small molecules and polymers can form helical aggregates through weak non-covalent interactions, while achiral Azo polymers can be induced to form chiral structures by chiral solvents or dopants. The review details the methods for constructing and regulating chiral helical structures, including the use of chiral limonene solvents, chiral dopants like tartaric acid, and photoisomerization of Azo units. These techniques enable the creation of stimuli-responsive chiroptical switches, such as "on-off," "amplification," and " inversion" switches, which are valuable for applications in chiral recognition, asymmetric catalysis, and chiral modulation. The review also explores the potential applications of these chiral nanomaterials, including in chiral recognition, asymmetric catalysis, chiral modulation, enantiomeric sensors, and liquid crystal (LC) materials. The dynamic control of chiral structures and their precise regulation are key challenges addressed in the review, with a focus on the development of smart-responsive multifunctional chiral nanomaterials. The future prospects of chiral nanomaterials are discussed, emphasizing the importance of further advancements in the field.This review discusses the construction, regulation, and application of chiral structures in azobenzene-containing systems. Chirality, a fundamental property in nature and biomolecules, plays a crucial role in various biological processes. The synthesis of chiral artificial structures is essential for understanding the origin of enantiomeric imbalance and its impact on biomolecular functions. Azobenzene (Azo) molecules, known for their reversible cis-trans isomerization, are widely used in functional dyes and materials due to their photoresponsive properties. The review highlights the dynamic regulation of chiroptical properties through temperature, solvent, magnetic field, and pH, enabling the construction of stimuli-responsive chiral nanomaterials with diverse functionalities. The construction of chiral structures in Azo-containing systems involves both chiral and achiral components. Chiral Azo small molecules and polymers can form helical aggregates through weak non-covalent interactions, while achiral Azo polymers can be induced to form chiral structures by chiral solvents or dopants. The review details the methods for constructing and regulating chiral helical structures, including the use of chiral limonene solvents, chiral dopants like tartaric acid, and photoisomerization of Azo units. These techniques enable the creation of stimuli-responsive chiroptical switches, such as "on-off," "amplification," and " inversion" switches, which are valuable for applications in chiral recognition, asymmetric catalysis, and chiral modulation. The review also explores the potential applications of these chiral nanomaterials, including in chiral recognition, asymmetric catalysis, chiral modulation, enantiomeric sensors, and liquid crystal (LC) materials. The dynamic control of chiral structures and their precise regulation are key challenges addressed in the review, with a focus on the development of smart-responsive multifunctional chiral nanomaterials. The future prospects of chiral nanomaterials are discussed, emphasizing the importance of further advancements in the field.