Citric Acid and the RNA World

Müller, Ulrich F.; Tor, Yitzhak

doi:10.1002/anie.201400847

Cited by 5 publications

(3 citation statements)

References 29 publications

(15 reference statements)

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“…Finally, the high concentrations of Na‐citrate may play an important role in segregating the ionizable lipid, and also may act as a reducing agent to suppress downstream mRNA degradation by quenching Mg 2+ ‐induced RNA fragmentation. [ 24 ]…”

Section: Discussionmentioning

confidence: 99%

“…Finally, the high concentrations of Na-citrate may play an important role in segregating the ionizable lipid, and also may act as a reducing agent to suppress downstream mRNA degradation by quenching Mg 2+ -induced RNA fragmentation. [24] The observation of bleb structures for LNP mRNA systems prepared in 25 mm NaOAc and containing second-generation lipids such as ALC-0315 (Figure 2f) suggests that optimization efforts aimed at developing ionizable lipids for improved transfection may well be optimizing their ability to promote bleb structure and thus enhanced mRNA stability rather than improved intracellular delivery. Ionizable lipids such as ALC-0315 and SM-102 exhibit pronounced "cone" shapes due to their branched acyl chain composition and would be expected to favor adoption of It is proposed that the presence of high concentrations of pH 4 buffers such as citrate causes fusion of the small vesicles and LNPs containing the mRNA initially produced on dilution of lipids in ethanol into an aqueous media.…”

Section: Discussionmentioning

confidence: 99%

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Induction of Bleb Structures in Lipid Nanoparticle Formulations of mRNA Leads to Improved Transfection Potency

Cheng,

Leung,

Zhang

et al. 2023

Advanced Materials

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The transfection potency of lipid nanoparticle (LNP) mRNA systems is critically dependent on the ionizable cationic lipid component. LNP mRNA systems composed of optimized ionizable lipids often display distinctive mRNA‐rich “bleb” structures. Here, it is shown that such structures can also be induced for LNPs containing nominally less active ionizable lipids by formulating them in the presence of high concentrations of pH 4 buffers such as sodium citrate, leading to improved transfection potencies both in vitro and in vivo. Induction of bleb structure and improved potency is dependent on the type of pH 4 buffer employed, with LNP mRNA systems prepared using 300 mm sodium citrate buffer displaying maximum transfection. The improved transfection potencies of LNP mRNA systems displaying bleb structure can be attributed, at least in part, to enhanced integrity of the encapsulated mRNA. It is concluded that enhanced transfection can be achieved by optimizing formulation parameters to improve mRNA stability and that optimization of ionizable lipids to achieve enhanced potency may well lead to improvements in mRNA integrity through formation of the bleb structure rather than enhanced intracellular delivery.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Induction of Bleb Structures in Lipid Nanoparticle Formulations of mRNA Leads to Improved Transfection Potency

Cheng,

Leung,

Zhang

et al. 2023

Advanced Materials

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“…Luckily, the coordination of Mg 2+ with citrate was recently shown to protect lipid vesicles while allowing non-enzymatic RNA polymerization to occur [ 93 ]. A similar mechanism may be possible to allow ribozyme-catalyzed RNA polymerization in lipid vesicles [ 94 ]. Another issue is the relatively low fidelity of template-dependent RNA polymerization, currently in the range of 97%, which may not be sufficient for the stable propagation of genetic information.…”

Section: Outlook: Outstanding Obstacles To Forging An Rna Worldmentioning

confidence: 99%

RNA Synthesis by in Vitro Selected Ribozymes for Recreating an RNA World

Martin

Unrau

Müller

2015

Life

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The RNA world hypothesis states that during an early stage of life, RNA molecules functioned as genome and as the only genome-encoded catalyst. This hypothesis is supported by several lines of evidence, one of which is the in vitro selection of catalytic RNAs (ribozymes) in the laboratory for a wide range of reactions that might have been used by RNA world organisms. This review focuses on three types of ribozymes that could have been involved in the synthesis of RNA, the core activity in the self-replication of RNA world organisms. These ribozyme classes catalyze nucleoside synthesis, triphosphorylation, and the polymerization of nucleoside triphosphates. The strengths and weaknesses regarding each ribozyme’s possible function in a self-replicating RNA network are described, together with the obstacles that need to be overcome before an RNA world organism can be generated in the laboratory.

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