Single-Particle Cryo-EM to Investigate Nonenveloped Virus-Like Particles

Nonenveloped viruses own numerous significant members that cause substantial agricultural losses and impact human health. Structural studies on these viruses are essential for understanding their mechanisms of pathogenesis and infection. Here, we describe protocols for determining high-resolution structures of nonenveloped viruses via cryogenic electron microscopy (cryo-EM). The steps include virus-like particle purification, cryo-grid preparation, cryo-EM image acquisition, and image processing for 3D reconstruction. These protocols can generally be applied to structural studies of all other nonenveloped viruses.