RNA Protection is Effectively Achieved by Pullulan Film Formation

Area of Science:

• Biochemistry
• Materials Science
• Molecular Biology

Background:

• Ribonucleic acid (RNA) is crucial for various biological functions but is prone to degradation by enzymes (RNases) and spontaneous chemical reactions.
• This instability limits RNA's long-term preservation and practical applications, especially in diagnostics and functional nucleic acid technologies.

Purpose of the Study:

• To develop a simple and effective method for preserving RNA integrity and function.
• To demonstrate the utility of this RNA stabilization technique in a practical biosensor application.

Main Methods:

• RNA stabilization was achieved by forming a protective film using pullulan, a polysaccharide derived from the fungus Aureobasidium pullulans.
• The pullulan film's protective effect against both spontaneous and RNase-catalyzed degradation was evaluated.
• A paper-based sensor utilizing a bacteria-detecting RNA-cleaving DNAzyme was engineered to showcase the method's practical utility.

Main Results:

• The pullulan film significantly inhibited both spontaneous and RNase-mediated degradation of RNA.
• The protective pullulan film was easily dissolved in aqueous solutions, allowing for the retrieval of fully functional RNA.
• A paper sensor incorporating the stabilized RNA-cleaving DNAzyme maintained its bacteria-detection capability after six months of storage at room temperature.

Conclusions:

• Pullulan film formation offers a straightforward and effective strategy for preserving RNA stability and function.
• This method enhances the potential for using RNA in various applications, including the development of robust and long-lasting diagnostic tools.
• The successful application in a paper-based bacterial detection sensor highlights the method's practical viability for field-deployable technologies.