Self-Assembly of Intracellular Multivalent RNA Complexes Using Dimeric Corn and Beetroot Aptamers.

Area of Science:

• Molecular Biology
• Biochemistry
• Synthetic Biology

Background:

• DNA and RNA can self-assemble into various structures.
• Current methods for self-assembling nucleic acids in mammalian cells are limited by instability and activation of innate RNA immunity.
• Genetically encoding self-assembly reactions in cells remains a challenge.

Purpose of the Study:

• To develop a method for inducing RNA self-assembly within living mammalian cells.
• To enable the creation of distinct RNA and RNA-protein assemblies.
• To overcome the limitations of existing self-assembly approaches.

Main Methods:

• Incorporation of dimeric fluorogenic RNA aptamers (Corn and Beetroot) into RNA constructs.
• Utilizing the self-dimerization properties of Corn and Beetroot aptamers to drive RNA-RNA interactions.
• Encoding distinct RNA-protein assemblies by leveraging the specificity of Corn and Beetroot homodimerization.

Main Results:

• Dimeric aptamers enable RNAs to self-assemble into large clusters within cells, mediated by multivalent RNA-RNA interactions.
• Corn and Beetroot aptamers form homodimers but not heterodimers, allowing for multiplexing.
• Proteins can be recruited to these self-assembled RNA structures, genetically encoding intracellular RNA-protein assemblies.

Conclusions:

• Dimeric aptamers provide a novel approach for inducing RNA self-assembly in mammalian cells.
• This method allows for the multiplexing of distinct RNA and RNA-protein assemblies within the same cell.
• The findings demonstrate the potential for genetically encoding intracellular RNA-protein assemblies through directed RNA self-assembly.