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Your search keyword '"Yoshinaga, Naoto"' showing total 38 results
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38 results on '"Yoshinaga, Naoto"'

1. Design of an Artificial Peptide Inspired by Transmembrane Mitochondrial Protein for Escorting Exogenous DNA into the Mitochondria to Restore their Functions by Simultaneous Multiple Gene Expression (Adv. Funct. Mater. 8/2024)

Author

Yoshinaga, Naoto, primary, Miyamoto, Takaaki, additional, Odahara, Masaki, additional, Takeda‐Kamiya, Noriko, additional, Toyooka, Kiminori, additional, Nara, Seia, additional, Nishimura, Haruna, additional, Ling, Feng, additional, Su'etsugu, Masayuki, additional, Yoshida, Minoru, additional, and Numata, Keiji, additional

Published
2024
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4. Design of an Artificial Peptide Inspired by Transmembrane Mitochondrial Protein for Escorting Exogenous DNA into the Mitochondria to Restore their Functions by Simultaneous Multiple Gene Expression

Author

Yoshinaga, Naoto, primary, Miyamoto, Takaaki, additional, Odahara, Masaki, additional, Takeda‐Kamiya, Noriko, additional, Toyooka, Kiminori, additional, Nara, Seia, additional, Nishimura, Haruna, additional, Ling, Feng, additional, Su'etsugu, Masayuki, additional, Yoshida, Minoru, additional, and Numata, Keiji, additional

Published
2023
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8. Phenylboronic Acid-Functionalized Polyplexes Tailored to Oral CRISPR Delivery

Author

Yoshinaga, Naoto, primary, Zhou, Joyce K., additional, Xu, Cong, additional, Quek, Chai Hoon, additional, Zhu, Yuefei, additional, Tang, Ding, additional, Hung, Lin Yung, additional, Najjar, Sarah A., additional, Shiu, Chin Ying Angela, additional, Margolis, Kara Gross, additional, Lao, Yeh-Hsing, additional, and Leong, Kam W., additional

Published
2023
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12. Bundling mRNA Strands to Prepare Nano-Assemblies with Enhanced Stability Towards RNase for In Vivo Delivery

Author

Yoshinaga, Naoto, Cho, Eol, Koji, Kyoko, Mochida, Yuki, Naito, Mitsuru, Osada, Kensuke, Kataoka, Kazunori, Cabral, Horacio, Uchida, Satoshi, and Kensuke, Osada

Abstract

Ribonuclease (RNase)‐mediated degradation of messenger RNA (mRNA) poses a huge obstruction to in vivo mRNA delivery. Herein, we propose a novel strategy to protect mRNA by structuring mRNA to prevent RNase attack through steric hinderance. Bundling of mRNA strands through hybridization of RNA oligonucleotide linkers allowed the preparation of mRNA nano‐assemblies (R‐NAs) comprised of 7.7 mRNA strands on average, mostly below 100 nm in diameter. R‐NA formation boosted RNase stability by around 100‐fold compared to naïve mRNA and preserved translational activity, allowing protein production. A mechanistic analysis suggests that an endogenous mRNA unwinding mechanism triggered by 5′‐cap‐dependent translation may induce selective R‐NA dissociation intracellularly, leading to smooth translation. R‐NAs showed efficient mRNA transfection in mouse brain, demonstrating the feasibility for in vivo administration.

Published
2019

13. Precise tuning of disulphide crosslinking in mRNA polyplex micelles for optimising extracellular and intracellular nuclease tolerability

Author

Dirisala, Anjaneyulu, Uchida, Satoshi, A Tockary, Theofilus, Yoshinaga, Naoto, Li, Junjie, Osawa, Shigehito, Gorantla, Lahari, Fukushima, Shigeto, Osada, Kensuke, Kataoka, Kazunori, and Kensuke, Osada

Subjects
technology, industry, and agriculture, macromolecular substances
Abstract

The major issues in messenger (m)RNA delivery are rapid mRNA degradation in the extracellular and intracellular spaces, which decreases the efficiency and duration for protein expression from mRNA. Stabilization of mRNA carriers using environment-responsive crosslinkings has promises to overcome these issues. Herein, we fine-tuned the structure of disulphide crosslinkings, which are selectively cleaved in the intracellular reductive environment, using the mRNA-loaded polyplex micelles (PMs) prepared from poly(ethylene glycol)-poly(L-lysine) (PEG-PLys) block copolymers, particularly by focussing on cationic charge density after the crosslinking. Primary amino groups in PLys segment were partially thiolated in two ways: One is to introduce 3-mercaptopropionyl (MP) groups via amide linkage, resulting in the decreased cationic charge density [PEG-PLys(MP)], and the other is the conversion of amino groups to 1-amidine-3-mercaptopropyl (AMP) groups with preserving cationic charge density [PEG-PLys(AMP)]. Compared to non-crosslinked and PEG-PLys(MP) PMs, PEG-PLys(AMP) PM attained tighter mRNA packaging in the PM core, thereby improving mRNA nuclease tolerability in serum and intracellular spaces, and providing enhanced protein expression in cultured cells at the optimal crosslinking density. These findings highlight the importance of cationic charge preservation in installing crosslinking moieties, providing a rationale for mRNA carrier design in the molecular level.

Published
2019

17. Trans influence of oxo and dithiolene coordination in oxidized models of molybdenum oxidoreductase: synthesis, structures, and properties of Q2 MoVIO2(1,2-benzenedithiolato)2 (Q = Net4, PPh4) and related complexes

Author

Ueyama, Norikazu, Oku, Hiroyuki, Kondo, Mitsuru, Okamura, Taka-aki, Yoshinaga, Naoto, and Nakamura, Akira

Subjects
Molybdenum compounds -- Research, Coordination compounds -- Research, Ligands -- Research, Chemistry
Abstract

The synthesis and characterization of three cis-dioxomolybdenum complexes containing dithione-like benzenedithiolate ligands, (PPh4)2 MoVIO2(1,2-benzenedithiolato)2 and (PPh4)2(MoVIO2 S2C2 CN sub2 sub2)2-, are presented. Structural analysis showed an octahedral structure with two long Mo-S bonds trans to each of the Mo=O groups and two short Mo-S bonds cis to them due to trans influence.

Published
1996

19. Phenylboronic Acid-Functionalized Micelles Dual-Targeting Boronic Acid Transporter and Polysaccharides for siRNA Delivery into Brown Algae

Author

Yoshinaga, Naoto, Miyamoto, Takaaki, Goto, Mami, Tanaka, Atsuko, and Numata, Keiji

Abstract

Brown algae play essential roles ecologically, practically, and evolutionarily because they maintain coastal areas, capture carbon dioxide, and produce valuable chemicals such as therapeutic drugs. To unlock their full potential, understanding the unique molecular biology of brown algae is imperative. Genetic engineering tools that regulate homeostasis in brown algae are essential for determining their biological mechanisms in detail. However, few methodologies have been developed to control gene expression due to the robust structural barriers of brown algae. To address this issue, we designed peptide-based, small interfering RNA (siRNA)-loaded micelles decorated with phenylboronic acid (PBA) ligands. The PBA ligands facilitated the cellular uptake of the micelles into a model brown alga, Ectocarpus siliculosus(E. Siliculosus), through chemical interaction with polysaccharides in the cell wall and biological recognition by boronic acid transporters on the plasma membrane. The micelles, featuring “kill two birds with one stone” ligands, effectively induced gene silencing related to auxin biosynthesis. As a result, the growth of E. siliculosuswas temporarily inhibited without persistent genome editing. This study demonstrated the potential for exploring the characteristics of brown algae through a simple yet effective approach and presented a feasible system for delivering siRNA in brown algae.

Published
2024
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20. Polyplex Micelles with Phenylboronate/Gluconamide Cross-Linking in the Core Exerting Promoted Gene Transfection through Spatiotemporal Responsivity to Intracellular pH and ATP Concentration

Author

Yoshinaga, Naoto, Ishii, Takehiko, Naito, Mitsuru, Endo, Taisuke, Uchida, Satoshi, Cabral, Horacio, Osada, Kensuke, and Kataoka, Kazunori

Abstract

Polyplexes as gene delivery carriers require integrated functionalities to modulate intracellular trafficking for efficient gene transfection. Herein, we developed plasmid DNA (pDNA)-loaded polyplex micelles (PMs) from poly(ethylene glycol)-based block catiomers derivatized with 4-carboxy-3-fluorophenylboronic acid (FPBA) and d-gluconamide to form pH- and ATP-responsive cross-linking in the core. These PMs exhibited robustness in the extracellular milieu and smooth endosomal escape after cellular uptake, and they facilitated pDNA decondensation triggered by increased ATP concentration inside of the cell. Laser confocal microscopic observation revealed that FPBA installation enhanced the endosomal escapability of the PMs; presumably, this effect resulted from the facilitated endo-/lysosomal membrane disruption triggered by the released block catiomers with hydrophobic FPBA moieties in the side chain from the PM at lower pH condition of endo-/lysosomes. Furthermore, the profile of intracellular pDNA decondensation from the PMs was monitored using Förster resonance energy transfer measurement by flow cytometry; these observations confirmed that PMs optimized for ATP-responsivity exerted effective intracellular decondensation of loaded pDNA to attain promoted gene transfection.

Published
2024
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24. Systemic Delivery of Folate-PEG siRNA Lipopolyplexes with Enhanced Intracellular Stability for In Vivo Gene Silencing in Leukemia

Author

Lee, Dian-Jang, primary, Kessel, Eva, additional, Lehto, Taavi, additional, Liu, Xueying, additional, Yoshinaga, Naoto, additional, Padari, Kärt, additional, Chen, Ying-Chen, additional, Kempter, Susanne, additional, Uchida, Satoshi, additional, Rädler, Joachim O., additional, Pooga, Margus, additional, Sheu, Ming-Thau, additional, Kataoka, Kazunori, additional, and Wagner, Ernst, additional

Published
2017
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25. Multilayered polyion complexes with dissolvable silica layer covered by controlling densities of cRGD-conjugated PEG chains for cancer-targeted siRNA delivery

Author

Naito, Mitsuru, primary, Azuma, Ryota, additional, Takemoto, Hiroyasu, additional, Hori, Mao, additional, Yoshinaga, Naoto, additional, Osawa, Shigehito, additional, Kamegawa, Rimpei, additional, Kim, Hyun Jin, additional, Ishii, Takehiko, additional, Nishiyama, Nobuhiro, additional, Miyata, Kanjiro, additional, and Kataoka, Kazunori, additional

Published
2017
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35. Bridging mRNA and Polycation Using RNA Oligonucleotide Derivatives Improves the Robustness of Polyplex Micelles for Efficient mRNA Delivery

Author

Yoshinaga, Naoto, Uchida, Satoshi, Dirisala, Anjaneyulu, Naito, Mitsuru, Koji, Kyoko, Osada, Kensuke, Cabral, Horacio, and Kataoka, Kazunori

Abstract

Polyplex for messenger RNA (mRNA) delivery requires strong yet reversible association between mRNA and polycation for extracellular robustness and selective intracellular disintegration. Herein, RNA oligonucleotide (OligoRNA) derivatives that bridge mRNA and polycation are developed to stabilize polyplex micelles (PMs). A set of the OligoRNAs introduced with a polyol moiety in their 5′ end is designed to hybridize to fixed positions along mRNA strand. After PM preparation from the hybridized mRNA and poly(ethylene glycol)‐polycation block copolymer derived with phenylboronic acid (PBA) moieties in its cationic segment, PBA moieties form reversible phenylboronate ester linkages with a polyol moiety at 5′ end of OligoRNAs and a diol moiety at their 3′ end ribose, in the PM core. The OligoRNAs work as a node to bridge ionically complexed mRNA and polycation, thereby improving PM stability against polyion exchange reaction and ribonuclease attack in extracellular environment. After cellular uptake, intracellular high concentration of adenosine triphosphate triggers the cleavage of phenylboronate ester linkages, resulting in mRNA release from PM. Ultimately, the PM provides efficient mRNA introduction in cultured cells and mouse lungs after intratracheal administration, demonstrating the potential of the bridging strategy in polyplex‐based mRNA delivery. Messenger RNA (mRNA) and polycation are bridged inside polyplex micelle (PM) using RNA oligonucleotide (OligoRNA) derivatives. OligoRNAs hybridize to mRNA via side chain complementary bases, and diol/polyol moieties at both ends of OligoRNAs bind to phenylboronic acid moieties installed in polycation. This strategy improves PM stability against nuclease and polyanions, thereby providing efficient mRNA introduction in vitro and in vivo.

Published
2022
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37. Comb-structured mRNA vaccine tethered with short double-stranded RNA adjuvants maximizes cellular immunity for cancer treatment.

Author

Tockary TA, Abbasi S, Matsui-Masai M, Hayashi A, Yoshinaga N, Boonstra E, Wang Z, Fukushima S, Kataoka K, and Uchida S

Subjects
Humans, Animals, Mice, Adjuvants, Immunologic pharmacology, Antigens, Immunity, Cellular, Cytokines genetics, RNA, Messenger genetics, Mice, Inbred C57BL, RNA, Double-Stranded genetics, Neoplasms therapy
Abstract

Integrating antigen-encoding mRNA (Messenger RNA) and immunostimulatory adjuvant into a single formulation is a promising approach to potentiating the efficacy of mRNA vaccines. Here, we developed a scheme based on RNA engineering to integrate adjuvancy directly into antigen-encoding mRNA strands without hampering the ability to express antigen proteins. Short double-stranded RNA (dsRNA) was designed to target retinoic acid-inducible gene-I (RIG-I), an innate immune receptor, for effective cancer vaccination and then tethered onto the mRNA strand via hybridization. Tuning the dsRNA structure and microenvironment by changing its length and sequence enabled the determination of the structure of dsRNA-tethered mRNA efficiently stimulating RIG-I. Eventually, the formulation loaded with dsRNA-tethered mRNA of the optimal structure effectively activated mouse and human dendritic cells and drove them to secrete a broad spectrum of proinflammatory cytokines without increasing the secretion of anti-inflammatory cytokines. Notably, the immunostimulating intensity was tunable by modulating the number of dsRNA along the mRNA strand, which prevents excessive immunostimulation. Versatility in the applicable formulation is a practical advantage of the dsRNA-tethered mRNA. Its formulation with three existing systems, i.e., anionic lipoplex, ionizable lipid-based lipid nanoparticles, and polyplex micelles, induced appreciable cellular immunity in the mice model. Of particular interest, dsRNA-tethered mRNA encoding ovalbumin (OVA) formulated in anionic lipoplex used in clinical trials exerted a significant therapeutic effect in the mouse lymphoma (E.G7-OVA) model. In conclusion, the system developed here provides a simple and robust platform to supply the desired intensity of immunostimulation in various formulations of mRNA cancer vaccines.

Published
2023
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38. Bundling mRNA Strands to Prepare Nano-Assemblies with Enhanced Stability Towards RNase for In Vivo Delivery.

Author

Yoshinaga N, Cho E, Koji K, Mochida Y, Naito M, Osada K, Kataoka K, Cabral H, and Uchida S

Subjects
Nucleic Acid Conformation, RNA Stability, Ribonucleases chemistry, Nanostructures chemistry, RNA, Messenger chemistry, Ribonucleases metabolism
Abstract

Ribonuclease (RNase)-mediated degradation of messenger RNA (mRNA) poses a huge obstruction to in vivo mRNA delivery. Herein, we propose a novel strategy to protect mRNA by structuring mRNA to prevent RNase attack through steric hinderance. Bundling of mRNA strands through hybridization of RNA oligonucleotide linkers allowed the preparation of mRNA nano-assemblies (R-NAs) comprised of 7.7 mRNA strands on average, mostly below 100 nm in diameter. R-NA formation boosted RNase stability by around 100-fold compared to naïve mRNA and preserved translational activity, allowing protein production. A mechanistic analysis suggests that an endogenous mRNA unwinding mechanism triggered by 5'-cap-dependent translation may induce selective R-NA dissociation intracellularly, leading to smooth translation. R-NAs showed efficient mRNA transfection in mouse brain, demonstrating the feasibility for in vivo administration., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Published
2019
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