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Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC

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Cross-Chiral, RNA-Catalyzed Exponential Amplification of RNA.

Grant A L Bare¹, Gerald F Joyce¹

¹The Salk Institute, 10010 North Torrey Pines Road, La Jolla, California 92037, United States.

Journal of the American Chemical Society

|November 3, 2021

Summary

This study demonstrates cross-chiral exponential amplification of RNA molecules, enabling the creation and evolution of RNA enzymes with opposite handedness. This breakthrough expands possibilities for prebiotic chemistry and the origins of life.

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Area of Science:

Biochemistry
Origin of Life Studies
Molecular Biology

Background:

Biological information macromolecules exhibit chiral uniformity (d-nucleotides in nucleic acids, l-amino acids in proteins).
This uniformity, while potentially expedient, is not a chemical imperative.
Previous research lacked methods to amplify RNA molecules of opposite chirality.

Purpose of the Study:

To demonstrate the feasibility of cross-chiral exponential amplification of RNA.
To amplify RNA molecules of the opposite handedness using an RNA enzyme.
To explore the potential for evolving RNA polymerases based on amplified RNA function.

Main Methods:

Utilized thermal cycling, similar to polymerase chain reaction (PCR).
Supplied oligonucleotide building blocks for both functional and complementary RNA strands.
Performed amplification in bulk solution and within water-in-oil emulsion droplets.

Main Results:

Achieved cross-chiral exponential amplification of RNA molecules.
Successfully amplified the l-RNA form of the hammerhead ribozyme using a d-RNA polymerase.
Demonstrated that the amplified l-hammerhead ribozyme possessed catalytic activity on an l-RNA substrate.
Showcased amplification within both bulk solution and compartmentalized emulsion systems.

Conclusions:

Cross-chiral exponential amplification is a viable method for generating RNA molecules of opposite handedness.
This technique allows for the amplification of functional RNA molecules, such as ribozymes.
The ability to amplify enantio-RNAs in various environments opens avenues for evolving novel RNA polymerases.