Recent advances in the analysis of the evolutionary RNA based therapeutics; a review.

The evolution of RNA-based therapies is bringing in a new era for the pharmaceutical industry. As a result of their crucial role in regulation of genes, the design of specific therapeutic oligonucleotides to bind to their RNA targets, making them promising candidates for targeted drug delivery and the treatment of various human diseases. RNA therapeutics are divided into different classes including antisense oligonucleotides (ASOs), RNA interference (RNAi), messenger RNA (mRNA), aptamers. Considering the complex nature RNA based therapies, the optimization of selective and sensitive analytical techniques for their quantification is crucial. These include liquid chromatography operating in various modes such as Ion Pair Reversed-Phase High-Performance Liquid Chromatography (IP-RP-HPLC), Anion Exchange Chromatography (AEC), Size exclusion chromatography (SEC) and Hydrophilic Interaction Liquid Chromatography (HILIC). Moreover, other techniques including capillary gel electrophoresis, and ELISA have been previously described for analysis of biological samples. In addition, a discussion of the vital role of Artificial Intelligence (AI) in the optimization of RNA therapeutics analysis in precision medicine is included. This work gives an in-depth analysis of the field of RNA therapeutics, including an overview of their various types, modes of action, applications, negative aspects, and examples of FDA-approved RNA-based drugs that are currently present in the market. Herein we highlight the various analytical procedures utilized for purification, characterization, and analysis of RNA therapies, which will be an invaluable guide for researchers to help them overcome the drawbacks in reported techniques allowing for the development of novel analytical methods that ensure accurate, and sensitive determination of RNA therapeutic oligonucleotides.