Lipid-peptide nanocomplexes for mRNA delivery in vitro and in vivo

Grant-Serroukh, Dania; Hunter, Morag R.; Maeshima, Ruhina; Tagalakis, Aristides D.; Aldossary, Ahmad M.; Allahham, Nour; Williams, Gareth R.; Edbrooke, Mark R.; Desai, Arpan; Hart, Stephen L.

doi:10.1016/j.jconrel.2022.06.018

Cited by 20 publications

(19 citation statements)

References 44 publications

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“…The inclusion of the DOPE in the liposomal formulation caused strong protein expression compared to DOPC or Chol. This result is similar to that of a previous report stating that mRNA lipoplexes containing DOPE exhibited higher transfection efficiency than those containing DOPC [31]. DOPE may improve gene expression by destabilizing the endosomal membrane after the cellular uptake of mRNA lipoplexes via endocytosis.…”

Section: Discussionsupporting

confidence: 91%

Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method

et al. 2023

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Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA complexes (siRNA lipoplexes) were prepared by mixing a lipid-ethanol solution with a siRNA solution. In this study, we applied the MEI method to prepare mRNA lipoplexes and evaluated the in vitro and in vivo protein expression efficiencies. We selected six cationic lipids and three neutral helper lipids to generate 18 mRNA lipoplexes. These were composed of cationic lipids, neutral helper lipids, and polyethylene glycol-cholesteryl ether (PEG-Chol). Among them, mRNA lipoplexes containing N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16) or 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl) propan-2-yl) amino)-N,N,N-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12) with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and PEG-Chol exhibited high protein expression in cells. Furthermore, mRNA lipoplexes composed of DC-1-16, DOPE, and PEG-Chol exhibited high protein expression in the lungs and spleen of mice after systemic injection and induced high antigen-specific IgG1 levels upon immunization. These results suggest that the MEI method can potentially increase the efficiency of mRNA transfection, both in vitro and in vivo.

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Section: Discussionsupporting

confidence: 91%

Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method

et al. 2023

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“…In the field of gene therapy, several authors have evaluated different lipid carriers for transfection processes. The use of a lipid-peptide nanocomplex that consists of a mixture of lipids and peptides that complex electrostatically with nucleic acids could be a delivery system for the sgRNA and Cas protein [47] that achieves better results than lipofectamine. Furthermore, other vehicles have been studied for the transfection of CRISPR/Cas9 RNP into cells for gene targeting, such as carbon quantum dots [48].…”

Section: Discussionmentioning

confidence: 99%

Production of Genetically Modified Porcine Embryos via Lipofection of Zona-Pellucida-Intact Oocytes Using the CRISPR/Cas9 System

Piñeiro-Silva

Navarro-Serna

Belda-Pérez

et al. 2023

Animals

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The generation of genetically modified pigs has an important impact thanks its applications in basic research, biomedicine, and meat production. Cloning was the first technique used for this production, although easier and cheaper methods were developed, such as the microinjection, electroporation, or lipofection of oocytes and zygotes. In this study, we analyzed the production of genetically modified embryos via lipofection of zona-pellucida-intact oocytes using LipofectamineTM CRISPRMAXTM Cas9 in comparison with the electroporation method. Two factors were evaluated: (i) the increment in the concentration of the lipofectamine–ribonucleoprotein complexes (LRNPC) (5% vs. 10%) and (ii) the concentration of ribonucleoprotein within the complexes (1xRNP vs. 2xRNP). We found that the increment in the concentration of the LRNPC had a detrimental effect on embryo development and a subsequent effect on the number of mutant embryos. The 5% group had a similar mutant blastocyst rate to the electroporation method (5.52% and 6.38%, respectively, p > 0.05). The increment in the concentration of the ribonucleoprotein inside the complexes had no effect on the blastocyst rate and mutation rate, with the mutant blastocyst rate being similar in both the 1xRNP and 2xRNP lipofection groups and the electroporation group (1.75%, 3.60%, and 3.57%, respectively, p > 0.05). Here, we showed that it is possible to produce knock-out embryos via lipofection of zona-pellucida-intact porcine oocytes with similar efficiencies as with electroporation, although more optimization is needed, mainly in terms of the use of more efficient vesicles for encapsulation with different compositions.

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“…The structure and size of the NA molecule affect binding. Therefore, plasmid DNA is a double-stranded circular molecule with a size ofapproximately 5-10 kb, whereas messenger RNA is linear and single-stranded, with varying sizes and secondary structures [4]. Moreover, because mRNA is much less stable and highly susceptible to RNase cleavage, it should be bound to a carrier immediately [5] or composed of modified nucleotides to increase its stability [6].…”

Section: Polyplexes As a Nucleic Acids Delivery System 21 Nucleic Aci...mentioning

confidence: 99%

Non-Viral Carriers for Nucleic Acids Delivery: Fundamentals and Current Applications

Shtykalova

Deviatkin

Freund

et al. 2023

Life

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Over the past decades, non-viral DNA and RNA delivery systems have been intensively studied as an alternative to viral vectors. Despite the most significant advantage over viruses, such as the lack of immunogenicity and cytotoxicity, the widespread use of non-viral carriers in clinical practice is still limited due to the insufficient efficacy associated with the difficulties of overcoming extracellular and intracellular barriers. Overcoming barriers by non-viral carriers is facilitated by their chemical structure, surface charge, as well as developed modifications. Currently, there are many different forms of non-viral carriers for various applications. This review aimed to summarize recent developments based on the essential requirements for non-viral carriers for gene therapy.

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Lipid-peptide nanocomplexes for mRNA delivery in vitro and in vivo

Cited by 20 publications

References 44 publications

Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method

Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method

Production of Genetically Modified Porcine Embryos via Lipofection of Zona-Pellucida-Intact Oocytes Using the CRISPR/Cas9 System

Non-Viral Carriers for Nucleic Acids Delivery: Fundamentals and Current Applications

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