4′-C-Trifluoromethyl modified oligodeoxynucleotides: synthesis, biochemical studies, and cellular uptake properties

2019 ◽  
Vol 17 (22) ◽  
pp. 5550-5560 ◽  
Author(s):  
Yifei Zhou ◽  
Chuanlong Zang ◽  
Huawei Wang ◽  
Jiajun Li ◽  
Zenghui Cui ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Cell Permeability ◽  
Biochemical Studies ◽  
Nuclease Resistance

Introducing a 4′-C-trifluoromethyl (4′-CF3) modification into oligodeoxynucleotides (ODNs) leads to improved nuclease resistance and increased cell permeability.

Lipophilic 2′-O-Acetal Ester RNAs: Synthesis, Thermal Duplex Stability, Nuclease Resistance, Cellular Uptake, and siRNA Activity after Spontaneous Naked Delivery

ChemBioChem ◽  
2016 ◽  
Vol 17 (21) ◽  
pp. 2054-2062 ◽  
Author(s):  
Annabelle Biscans ◽  
Jean-Rémi Bertrand ◽  
Josephine Dubois ◽  
Jacqueline Rüger ◽  
Jean-Jacques Vasseur ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Duplex Stability ◽  
Nuclease Resistance

scholarly journals Shaping Rolling Circle Amplification Products into DNA Nanoparticles by Incorporation of Modified Nucleotides and Their Application to In Vitro and In Vivo Delivery of a Photosensitizer

Molecules ◽  
2018 ◽  
Vol 23 (7) ◽  
pp. 1833 ◽  
Author(s):  
Kyoung-Ran Kim ◽  
Pascal Röthlisberger ◽  
Seong Kang ◽  
Kihwan Nam ◽  
Sangyoup Lee ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Cellular Uptake ◽  
Rolling Circle Amplification ◽  
Modified Nucleotides ◽  
Rolling Circle ◽  
Dna Nanoparticles ◽  
Nuclease Resistance ◽  
Cellular Uptake Efficiency

Rolling circle amplification (RCA) is a robust way to generate DNA constructs, which are promising materials for biomedical applications including drug delivery because of their high biocompatibility. To be employed as a drug delivery platform, however, the DNA materials produced by RCA need to be shaped into nanoparticles that display both high cellular uptake efficiency and nuclease resistance. Here, we showed that the DNA nanoparticles (DNPs) can be prepared with RCA and modified nucleotides that have side-chains appended on the nucleobase are capable of interacting with the DNA strands of the resulting RCA products. The incorporation of the modified nucleotides improved cellular uptake efficiency and nuclease resistance of the DNPs. We also demonstrated that these DNPs could be employed as carriers for the delivery of a photosensitizer into cancer cells to achieve photodynamic therapy upon irradiation at both the in vitro and in vivo levels.

Synthesis, binding, nuclease resistance and cellular uptake properties of 2′- O -acetalester-modified oligonucleotides containing cationic groups

2015 ◽  
Vol 23 (17) ◽  
pp. 5360-5368 ◽  
Author(s):  
Annabelle Biscans ◽  
Sonia Rouanet ◽  
Jean-Rémi Bertrand ◽  
Jean-Jacques Vasseur ◽  
Christelle Dupouy ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Modified Oligonucleotides ◽  
Nuclease Resistance

scholarly journals Unprotected Peptide Macrocyclization and Stapling via A Fluorine-Thiol Displacement Reaction

2020 ◽  
Author(s):  
Md Shafiqul Islam ◽  
Samuel L. Junod ◽  
Si Zhang ◽  
Zakey Yusuf Buuh ◽  
Yifu Guan ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Protein Interactions ◽  
Cell Permeability ◽  
Displacement Reaction ◽  
Protein Protein Interactions ◽  
Cancer Cell Growth ◽  
Stapled Peptides ◽  
Peptide Substrates ◽  
New Approach ◽  
Dynamics Simulations

AbstractStapled peptides serve as a powerful tool for probing protein-protein interactions, but its application has been largely impeded by the limited cellular uptake. Here we report the discovery of a facile peptide macrocyclization and stapling strategy based on a fluorine thiol displacement reaction (FTDR), which renders a class of peptide analogues with enhanced stability, affinity, and cell permeability. This new approach enabled selective modification of the orthogonal fluoroacetamide side chains in unprotected peptides, with the identified 1,3-benzenedimethanethiol linker promoting alpha helicity of a variety of peptide substrates, as corroborated by molecular dynamics simulations. The cellular uptake of these stapled peptides was universally enhanced compared to the classic ring-closing metathesis (RCM) stapled peptides. Pilot mechanism studies suggested that the uptake of FTDR-stapled peptides may involve multiple endocytosis pathways. Consistent with the improved cell permeability, the FTDR-stapled lead Axin analogues demonstrated better inhibition of cancer cell growth than the RCM-stapled analogues.Graphical Abstract

scholarly journals Biocompatibility Profile and In Vitro Cellular Uptake of Self-assembled Alginate Nanoparticles

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 555 ◽  
Author(s):  
Pei Zhang ◽  
Shirui Zhao ◽  
Yaoyao Yu ◽  
Huan Wang ◽  
Yan Yang ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Polymeric Nanoparticles ◽  
Cell Permeability ◽  
Intestinal Epithelial ◽  
Cellular Delivery ◽  
Concentration Time ◽  
Self Assembled ◽  
Uptake Studies ◽  
Cellular Uptake Efficiency

Polymeric nanoparticles could offer promising controlled drug delivery. The biocompatibility is of extreme importance for future applications in humans. Self-assembled polymeric nanoparticles based on phenylalanine ethyl ester (PAE)-modified alginate (Alg) had been successfully prepared and characterized in our lab. However, little is known about their interaction with cells and other biological systems. In this study, nanoparticles (NPs) based on PAE-Alg conjugates (PEA-NPs) with different degree of substitution (DS) were prepared and investigated. Our results showed that PEA-NPs had no effects on the proliferation of the human intestinal epithelial Caco-2 cells at concentrations up to 1000 μg/mL. Furthermore, the in vitro cellular uptake profile of PEA-NPs, concerning several parameters involved in the application of therapeutic or diagnostic NPs, such as NPs concentration, time and temperature, was described. Different NPs have been adopted for cellular uptake studies and the NPs internalized into Caco-2 cells were quantified. Cellular uptake efficiency could reach 60% within 4 h. PEA-NPs also showed greater cell permeability than oleoyl alginate ester nanoparticles (OAE-NPs) previously prepared in our lab. Our studies reveal that NPs based on PEA conjugate are promising nanosystems for cellular delivery.

scholarly journals A critical analysis of methods used to investigate the cellular uptake and subcellular localization of RNA therapeutics

2020 ◽  
Vol 48 (14) ◽  
pp. 7623-7639 ◽  
Author(s):  
Kirsten Deprey ◽  
Nefeli Batistatou ◽  
Joshua A Kritzer
Keyword(s):  
Cellular Uptake ◽  
Critical Analysis ◽  
Genetic Diseases ◽  
Specific Protein ◽  
Cell Penetration ◽  
Chemical Modifications ◽  
Aberrant Splicing ◽  
Promising Strategy ◽  
Nuclease Resistance ◽  
Delivery Strategies

Abstract RNA therapeutics are a promising strategy to treat genetic diseases caused by the overexpression or aberrant splicing of a specific protein. The field has seen major strides in the clinical efficacy of this class of molecules, largely due to chemical modifications and delivery strategies that improve nuclease resistance and enhance cell penetration. However, a major obstacle in the development of RNA therapeutics continues to be the imprecise, difficult, and often problematic nature of most methods used to measure cell penetration. Here, we review these methods and clearly distinguish between those that measure total cellular uptake of RNA therapeutics, which includes both productive and non-productive uptake, and those that measure cytosolic/nuclear penetration, which represents only productive uptake. We critically analyze the benefits and drawbacks of each method. Finally, we use key examples to illustrate how, despite rigorous experimentation and proper controls, our understanding of the mechanism of gymnotic uptake of RNA therapeutics remains limited by the methods commonly used to analyze RNA delivery.

scholarly journals Preparation of siRNA–PLGA/Fabʹ–PLGA mixed micellar system with target cell-specific recognition

2021 ◽  
Author(s):  
Mai Hazekawa ◽  
Takuya Nishinakagawa ◽  
Takeshi Mori ◽  
Miyako Yoshida ◽  
Takahiro Uchida ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Target Cell ◽  
Cancer Metastasis ◽  
Mixed Micelles ◽  
Sirna Delivery ◽  
Cell Permeability ◽  
Target Cells ◽  
Small Interfering Rnas

Abstract Small interfering RNAs (siRNAs) are susceptible to nucleases and degrade quickly in vivo. Moreover, siRNAs demonstrate poor cellular uptake and cannot cross the cell membrane because of its polyanionic characteristics. To overcome these challenges, an intelligent gene delivery system that protects siRNAs from nucleases and facilitates siRNA cellular uptake is required. We previously reported the potential of siRNA-poly(D,L-lactic-co-glycolic acid; PLGA) micelles as an effective siRNA delivery tool in a murine peritoneal dissemination model by local injection. However, there was no effective formulation for siRNA delivery to target cells via intravenous injection. This study aimed to prepare siRNA–PLGA/Fabʹ–PLGA mixed micelles for siRNA delivery to target floating cells and evaluate its formulation in vitro. As the target siRNA protein in CEMx174, CyclinB1 levels were significantly reduced when siRNA–PLGA/Fabʹ–PLGA mixed micelles were added to cells compared with siRNA–PLGA micelles. siRNA–PLGA/Fabʹ–PLGA mixed micelles have high cell permeability and high target cell accumulation by endocytosis because flow cytometry detected labeling micelles in target cells. This study supports siRNA–PLGA/Fabʹ–PLGA mixed micelles as an effective siRNA delivery tool. This formulation can be administered systemically in dosage form against target cells, including cancer metastasis or blood cancer.

scholarly journals Preparation of siRNA–PLGA/Fabʹ–PLGA mixed micellar system with target cell-specific recognition

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mai Hazekawa ◽  
Takuya Nishinakagawa ◽  
Takeshi Mori ◽  
Miyako Yoshida ◽  
Takahiro Uchida ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Target Cell ◽  
Cancer Metastasis ◽  
Mixed Micelles ◽  
Sirna Delivery ◽  
Cell Permeability ◽  
Target Cells ◽  
Small Interfering Rnas

AbstractSmall interfering RNAs (siRNAs) are susceptible to nucleases and degrade quickly in vivo. Moreover, siRNAs demonstrate poor cellular uptake and cannot cross the cell membrane because of its polyanionic characteristics. To overcome these challenges, an intelligent gene delivery system that protects siRNAs from nucleases and facilitates siRNA cellular uptake is required. We previously reported the potential of siRNA-poly(d,l-lactic-co-glycolic acid; PLGA) micelles as an effective siRNA delivery tool in a murine peritoneal dissemination model by local injection. However, there was no effective formulation for siRNA delivery to target cells via intravenous injection. This study aimed to prepare siRNA–PLGA/Fabʹ–PLGA mixed micelles for siRNA delivery to target floating cells and evaluate its formulation in vitro. As the target siRNA protein in CEMx174, CyclinB1 levels were significantly reduced when siRNA–PLGA/Fabʹ–PLGA mixed micelles were added to cells compared with siRNA–PLGA micelles. siRNA–PLGA/Fabʹ–PLGA mixed micelles have high cell permeability and high target cell accumulation by endocytosis because flow cytometry detected labeling micelles in target cells. This study supports siRNA–PLGA/Fabʹ–PLGA mixed micelles as an effective siRNA delivery tool. This formulation can be administered systemically in dosage form against target cells, including cancer metastasis or blood cancer.

Ruthenium Red Staining of Human Hard Keratin Fibers

1982 ◽  
Vol 40 ◽  
pp. 278-279
Author(s):  
M. C. Buhrer ◽  
R. A. Mathews
Keyword(s):  
Human Hair ◽  
Ruthenium Red ◽  
Acid Mucopolysaccharides ◽  
Biochemical Studies ◽  
Keratin Fibers ◽  
Ruthenium Red Staining ◽  
Extracellular Materials

Ruthenium red has been used as a stain to demonstrate a variety of extracellular materials, especially acid mucopolysaccharides. It also reacts with certain intracellular and extracellular lipids. Since biochemical studies in our laboratory demonstrated the presence of a variety of monosaccharides in human hair ruthenium red staining procedures were adopted in order to evaluate the presence and morphological location of acid oligosaccharides in the keratinized aspect of hair.

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