Methods and advances in RNA segment-selective labelling
image: PLOR-based RNA segmental labeling. PLOR consists of three stages: initiation, extension, and termination. Transcription occurs on a DNA template immobilized on beads. In a single cycle of the initiation or extension phase, three or fewer rNTPs are added, causing transcription to pause due to the absence of the required rNTP. After removing residual rNTPs and introducing labeled rNTPs, extension resumes, generating segment-labeled RNA
Credit: HIGHER EDUCATION PRESS
RNA is fundamental to all life, governing gene expression, regulating biological processes, and influencing growth, development, and cellular metabolism. However, elucidating the precise mechanisms of these functions requires methods to probe the complex structure, dynamics, and interactions of RNA molecules. In recent years, RNA segment-selective labeling has emerged as an essential technique for studying RNA structure, function, and dynamics. We classify various RNA segmental labeling methods based on their underlying principles into three categories: chemical methods (including solid-phase synthesis labeling and post-synthesis RNA chemical ligation), ligase-based methods (such as T4 RNA/DNA ligases, deoxyribozymes, and ribozymes), and polymerase-based methods (by DNA polymerase and T7 RNA polymerase). Each method has its unique characteristics and needs to be selected according to specific experimental requirements. Despite significant progress, current methods still face challenges. This review focuses on RNA segment-selective labeling techniques, exploring their underlying principles, advantages, limitations, and applications. The aim is to provide a foundation for developing novel RNA segmental labeling techniques with broader application potential in the future.
The work entitled “Methods and advances in RNA segment-selective labelling” was published in Biophysics Reports (ahead of print in 2026).