In this study, we examined how the regioregularity of poly(3-hexylthiophene) (P3HT) affects molecular packing, free volume, charge transport, and gas sensing properties. Our results showed that the presence of regular alkyl side chains on the polymer backbone promoted a high degree of structural order in regioregular P3HT molecules, leading to a compact packing density and reduced free volume. Consequently, it was more challenging for NO2 molecules to interact with the hole charge carriers in the conductive channel. On the other hand, the regiorandom P3HT films displayed a larger free volume, attributed to the irregular side chains, which facilitated the gas-analyte interaction while impeding efficient charge transport. Thus, these films exhibited greater sensitivity to analyte gas molecules. The molecular order, packing density, and hardness of P3HT films were confirmed through the use of multiple techniques, including UV-vis spectroscopy, atomic force microscopy, and grazing-incidence X-ray diffraction. Additionally, the regiorandom P3HT films showed enhanced mechanical flexibility compared to the regioregular films. In conclusion, our findings emphasize that the regularity of polymer molecules plays a significant role in determining the charge carrier transport and gas adsorption characteristics.