Spacer Fidelity Assessments of Guide RNA by Top-Down Mass Spectrometry.

ABSTRACT: The advancement of CRISPR-based gene editing tools into biotherapeutics offers the potential for cures to genetic disorders and for new treatment paradigms for even common diseases. Arguably, the most important component of a CRISPR-based medicine is the guide RNA, which is generally large (>100-mer) synthetic RNA composed of a "tracr" and "spacer" region, the latter of which dictates the on-target editing site as well as potential undesired off-target edits. Aiming to advance contemporary capabilities for gRNA characterization to ensure the spacer region is of high fidelity, top-down mass spectrometry was herein implemented to provide direct and quantitative assessments of highly modified gRNA. In addition to sequencing the spacer region and pinpointing modifications, top-down mass spectr

SUBMITTER: Macias LA 

PROVIDER: S-EPMC10375574 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature