Abstarct: Abstract Significant advancements in molecular dynamics simulation and nanotechnology over the past decades have established nanocarriers as a crucial and widely applied topic in life sciences and engineering research. DNA origami nanotubes, due to their unique mechanical and chemical properties and versatile capabilities, hold great potential in nanotechnology and biomedicine. Given the importance of nanocarriers in molecular robotics and the need to ensure structural integrity and bond stability under various environmental conditions and vibrations, understanding the dynamic behavior and vibrational analysis of these nanocarriers is of particular significance. In this study, the dynamic behavior and vibrational analysis of DNA origami nanotubes under different environmental conditions were investigated using steered molecular dynamics (SMD) simulations and an analytical mechanical model. First, a DNA origami nanotube was designed using the caDNAno software, and SMD simulations were performed with GROMACS to evaluate its elasticity at different temperatures under a constant stretching rate. Further simulations were conducted at various stretching rates at a fixed temperature. To assess the structural changes and flexibility of the DNA origami nanotube under these conditions, after energy optimization, the Young's modulus was calculated and analyzed using GROMACS. Subsequently, to accelerate the natural frequency analysis, the Euler–Bernoulli beam (EBNB) model was employed. The Young’s modulus values were incorporated into the Euler–Bernoulli beam equation, which was solved using the transfer matrix method under different boundary conditions, temperature variations, and stretching rates. The molecular dynamics simulation results revealed structural changes in the DNA origami nanotube across different temperatures and stretching rates. The Young's modulus gradually decreased with increasing temperature, indicating greater flexibility of the nanotube, while it increased with higher stretching rates. The results from solving the Euler–Bernoulli beam equation showed that rising temperatures increased the nanotube's natural frequencies, whereas higher stretching rates tended to decrease them. Validation with precise solutions confirmed the accuracy of the transfer matrix method in predicting the dynamic behavior of DNA nanostructures. This study provides valuable insights into the mechanical and vibrational properties of DNA origami nanotubes, paving the way for further advancements in nanometric engineering and DNA-baxsed technologies.