Cell-internalization SELEX: method for identifying cell-internalizing RNA aptamers for delivering siRNAs to target...

Area of Science:

• Biotechnology
• Molecular Biology
• Drug Delivery

Background:

• Cellular uptake remains a key challenge for RNA interference (RNAi) therapeutics.
• Recent clinical trials show promise for RNAi drugs targeting liver diseases, demonstrating safe and effective gene knockdown.
• Expanding RNAi therapy to other organs requires improved delivery systems, such as conjugating small interfering RNAs (siRNAs) to RNA aptamers.

Purpose of the Study:

• To develop a rapid method for identifying RNA aptamers capable of delivering siRNA drugs into the cytoplasm of diverse cell types.
• To overcome the limitations of current delivery techniques for RNAi-based therapeutics.

Main Methods:

• A novel cell-based selection process termed "cell-internalization SELEX" (Systematic Evolution of Ligands by Exponential Enrichment) was employed.
• This process integrates cell culture-based SELEX, next-generation sequencing (NGS), and advanced bioinformatics tools.
• The method focuses on identifying RNA aptamer sequences that facilitate cellular entry and cytoplasmic delivery.

Main Results:

• The study describes a process for the rapid identification and characterization of RNA aptamers.
• These aptamers are specifically selected for their ability to deliver siRNA into target cells.
• The developed method aims to enable the use of aptamer-siRNA conjugates for broader therapeutic applications.

Conclusions:

• The cell-internalization SELEX process offers a viable solution for discovering RNA aptamers for siRNA delivery.
• This advancement is critical for developing RNAi therapeutics for diseases affecting various organ systems.
• The combination of SELEX, NGS, and bioinformatics accelerates the identification of effective RNA delivery tools.