Room-temperature manipulation and processing of information encoded in the electronic valley pseudospin and spin degrees of freedoms lie at the heart of the next technological quantum revolution. In atomically thin layers of transition-metal dichalcogenides (TMDs) with hexagonal lattices, valley-polarized excitations and valley quantum coherence can be generated by simply shining with adequately polarized light. In turn, the polarization states of light can induce topological Hall currents in the absence of an external magnetic field, which underlies the fundamental principle of opto-valleytronics devices. However, demonstration of optical generation of valley polarization at room temperature has remained challenging and not well understood. Here, we demonstrate control of strong valley polarization (valley quantum coherence) at room temperature of up to ∼50% (∼20%) by strategically designing Coulomb forces and spin−orbit interactions in atomically thin TMDs via chalcogenide alloying. We show that tailor making the carrier density and the relative order between optically active (bright) and forbidden (dark) states by key variations on the chalcogenide atom ratio allows full control of valley pseudospin dynamics. Our findings set a comprehensive approach for intrinsic and efficient manipulation of valley pseudospin and spin degree of freedom toward realistic opto-valleytronics devices.