Towards the design and construction of self-replicating RNA nanostructures
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Date
2024
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Abstract
The transition from an RNA-based world to DNA as the primary genetic material is a
pivotal topic in origin-of-life research. This study aims to prototype a hybrid genome
system combining DNA and RNA, capable of self-replication and evolution, initially in
vitro and potentially in vivo. Using Qß replicase, segmented RNA genomes were
synthesized, incorporating the Phi29 phage pRNA for self-assembly into nanorings. The
replication efficacy of these replicons was evaluated both independently and in coupled
assembly-replication reactions, with Qß chosen for its high amplification efficiency and
template versatility.
To mitigate the challenge of parasite RNA formation, which depletes essential replication
components, a water-emulsion system was employed. The modular assembly of
replicons into nanorings was further investigated, with a focus on system reproducibility
and the effect of Mg2+ concentrations on nanoring stability. The final goal is to integrate
self-replication with self-assembly, thereby creating a segmented, modular RNA genome
for in vitro genetic information storage. This work addresses key challenges and
advancements in developing synthetic self-replication systems for eventual genomic
transplantation in E. coli.
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Keywords
RNA, Self-replicating RNA, Cell-free, pRNA