Polymeric Micelles for Messenger RNA Delivery: Synthesis, Characterization, and In Vitro Release

• Grant number: 25/22904-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2026
• End date: January 31, 2027
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Physical-Chemistry
• Principal Investigator: Marcio José Tiera
• Grantee: Ana Laura Ferraz Tezoto
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
• Associated research grant: 23/03182-4 - Polymeric Nanoparticles for Gene Therapy: Development of vectors for delivery of messenger RNA and interfering RNA, AP.R

Abstract

The delivery of nucleic acids, such as mRNA and siRNA, represents a promising therapeutic approach for vaccines, cancer immunotherapy, and the treatment of genetic disorders. However, the clinical application of these biopolymers requires the development of efficient carriers capable of promoting nucleic acid complexation, protection, and intracellular release. This project proposes the synthesis of amphiphilic chitosan derivatives modified with tertiary amine groups and hydrophobic cholesterol units, aiming to optimize the formation of stable nanoparticles responsive to endosomal pH. Samples will be characterized by NMR, IR, gel permeation chromatography, potentiometric titration, and dynamic light scattering, in addition to the determination of the critical association concentration (CAC). The efficiency of siRNA and mRNA complexation will be assessed by agarose gel electrophoresis, followed by cytotoxicity assays and in vitro transfection studies in HeLa cells. It is expected that polymer compositions balancing electrostatic and hydrophobic interactions will be identified, ensuring high colloidal stability and transfection efficiency with low toxicity. The results will contribute to the advancement of polymeric systems as non-viral vectors, offering an alternative to conventional lipid nanoparticles (AU)