» Articles » PMID: 39345227

Analytical Methods to Evaluate RNA Circularization Efficiency

Overview
Journal Electrophoresis
Specialty Chemistry
Date 2024 Sep 30
PMID 39345227
Authors
Affiliations
Soon will be listed here.
Abstract

Circular RNAs (circRNAs) have emerged as pivotal players in RNA therapeutics. Unlike linear counterparts, circRNAs possess a closed-loop structure, conferring them with enhanced stability and resistance to degradation. Ribozyme-based strategy stands out as the predominant method for synthetic circRNA production, by precisely cleaving and promoting the formation of a covalent circular structure. However, there is still a lack of analytical methods that can provide high-throughput and quantitative analysis to facilitate the circRNA vector engineering process. In the report, we detail analytical methods to characterize and evaluate ribozyme-based RNA circularization efficiency. Our approach will capture the attention of researchers interested in optimizing RNA circularization efficiency, as well as those focused on exploring key elements for ribozyme catalytic activity.

References
1.
Puttaraju M, Been M . Group I permuted intron-exon (PIE) sequences self-splice to produce circular exons. Nucleic Acids Res. 1992; 20(20):5357-64. PMC: 334342. DOI: 10.1093/nar/20.20.5357. View

2.
Verduci L, Tarcitano E, Strano S, Yarden Y, Blandino G . CircRNAs: role in human diseases and potential use as biomarkers. Cell Death Dis. 2021; 12(5):468. PMC: 8113373. DOI: 10.1038/s41419-021-03743-3. View

3.
Zhou W, Cai Z, Liu J, Wang D, Ju H, Xu R . Circular RNA: metabolism, functions and interactions with proteins. Mol Cancer. 2020; 19(1):172. PMC: 7734744. DOI: 10.1186/s12943-020-01286-3. View

4.
Barbier A, Jiang A, Zhang P, Wooster R, Anderson D . The clinical progress of mRNA vaccines and immunotherapies. Nat Biotechnol. 2022; 40(6):840-854. DOI: 10.1038/s41587-022-01294-2. View

5.
Baden L, El Sahly H, Essink B, Kotloff K, Frey S, Novak R . Efficacy and Safety of the mRNA-1273 SARS-CoV-2 Vaccine. N Engl J Med. 2020; 384(5):403-416. PMC: 7787219. DOI: 10.1056/NEJMoa2035389. View