Guide RNAs in CRISPR-based therapeutics: addressing CMC and analytical challenges to improve clinical translation

“[...] there is a need to identify the key next steps that the industry must take over the coming years to ensure that analytical frameworks keep pace with the increasing complexity of these therapeutics.”On December 19, 2025, Róisin McGuigan, Editor, Nucleic Acid Insights, spoke to Kok-Seong Lim, Independent Expert in Cell and Gene Therapy Chemistry, Manufacturing and Control (CMC), about the role of guide RNAs (gRNAs) in CRISPR-based therapeutics, focusing on starting material choices, gRNA-specific CMC and analytical challenges, regulatory priorities, and emerging approaches needed to support successful clinical translation. This article has been written based on that interview.Robust CMC strategies and fit-for-purpose analytical frameworks are essential to the successful development of guide RNA for CRISPR-based therapeutics. In this article, Kok-Seong Lim explores how starting and raw material selection, manufacturing processes and analytical methods evolve from research settings to late-phase development and commercial supply. He discusses how oligonucleotide length and impurity profiles differentiate guide RNAs from traditional oligonucleotide modalities and highlights current analytical limitations. He also addresses regulatory priorities, including regulatory harmonization, the application of existing guidance and the qualification of emerging technologies, and considers how platform approaches and novel manufacturing strategies may enable scalable and robust development of CRISPR-based therapeutics.