This paper presents the novel development of the dynamic stiffness matrix for a three-layered symmetric pre-twisted sandwich beam (PTSB), aiming to investigate its free vibration characteristics. The outer layers of the beam are modeled using the Euler–Bernoulli theory, while the core is assumed to deform solely in shear. The boundary conditions and governing partial differential equations of motion are derived based on Hamilton's principle. By applying harmonic variations of displacements, the governing equations of motion are expressed as a tenth-order equation, which is solved to obtain the desired dynamic stiffness matrix. To compute the natural frequencies of in-plane and out-of-plane free vibration for both uniform and PTSBs, the Wittrick–Williams algorithm is employed. The computed frequencies are compared with the results obtained by other authors as well as those obtained from ABAQUS simulations. Various vibration modes of uniform and twisted sandwich beams are plotted and thoroughly discussed. Interestingly, contrary to straight symmetric sandwich beams, the results indicate that flexural displacements in pre-twisted symmetric sandwich beams exhibit coupling in two planes. Additionally, although there are minor changes in vibration frequencies, the mode shapes undergo significant transformations as the pre-twist angle is altered.