Cell-penetrating poly(disulfide)s-based nanoquenchers (qCPDs) for self-monitoring of intracellular gene delivery

Appropriate gene delivery systems are essential for successful gene therapy in clinical medicine. Lipid-mediated nucleic acid delivery is an alternative to viral vector-mediated gene delivery and has the following advantages. Lipid-mediated delivery of DNA or mRNA is usually more rapid than viral-mediated delivery, offers a larger payload, and has a nearly zero risk of incorporation. Lipid-mediated delivery of DNA or RNA is therefore preferable to viral DNA delivery in those clinical applications that do not require long-term expression for chronic conditions. Delivery of RNA may be preferable to non-viral DNA delivery in some clinical applications, since transit across the nuclear membrane is not necessary, and onset of expression with RNA is therefore even faster than with DNA, although both are faster than most viral vectors. Delivery of RNA to target organ(s) has previously been challenging due to RNA’s rapid degradation in biological systems, but cationic lipids complexed with RNA, as well as lipid nanoparticles (LNPs), have allowed for delivery and expression of the complexed RNA both in vitro and in vivo. This review will focus on the non-viral lipid-mediated delivery of RNAs, including mRNA, siRNA, shRNA, and microRNA, to the central nervous system (CNS), an organ with at least two unique challenges. The CNS contains a large number of slowly dividing or non-dividing cell types and is protected by the blood brain barrier (BBB). In non-dividing cells, RNA-lipid complexes demonstrated increased transfection efficiency relative to DNA transfection. The efficiency, timing of the onset, and duration of expression after transfection may determine which nucleic acid is best for which proposed therapy. Expression can be seen as soon as 1 h after RNA delivery, but duration of expression has been limited to 5–7 h. In contrast, transfection with a DNA lipoplex demonstrates protein expression within 5 h and lasts as long as several weeks after transfection.

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Development and Characterization of High Efficacy Cell-Penetrating Peptide via Modulation of the Histidine and Arginine Ratio for Gene Therapy

Materials ◽

10.3390/ma14164674 ◽

2021 ◽

Vol 14 (16) ◽

pp. 4674

Author(s):

Yu Liu ◽

Huan-Huan Wan ◽

Duo-Mei Tian ◽

Xiao-Jun Xu ◽

Chang-Long Bi ◽

...

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Cell Penetrating Peptides ◽

Viral Gene ◽

Main Role ◽

Delivery Vector ◽

Gene Delivery Vectors ◽

Transfection Efficacy ◽

Cell Penetrating ◽

Viral Gene Delivery

Cell-penetrating peptides (CPPs), as non-viral gene delivery vectors, are considered with lower immunogenic response, and safer and higher gene capacity than viral systems. In our previous study, a CPP peptide called RALA (arginine rich) presented desirable transfection efficacy and owns a potential clinic use. It is believed that histidine could enhance the endosome escaping ability of CPPs, yet RALA peptide contains only one histidine in each chain. In order to develop novel superior CPPs, by using RALA as a model, we designed a series of peptides named HALA (increased histidine ratio). Both plasmid DNA (pDNA) and siRNA transfection results on three cell lines revealed that the transfection efficacy is better when histidine replacements were on the C-terminal instead of on the N-terminal, and two histidine replacements are superior to three. By investigating the mechanism of endocytosis of the pDNA nanocomplexes, we discovered that there were multiple pathways that led to the process and caveolae played the main role. During the screening, we discovered a novel peptide-HALA2 of high cellular transfection efficacy, which may act as an exciting gene delivery vector for gene therapy. Our findings also bring new insights on the development of novel robust CPPs.

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Chitosan-Based Drug Delivery System: Applications in Fish Biotechnology

Polymers ◽

10.3390/polym12051177 ◽

2020 ◽

Vol 12 (5) ◽

pp. 1177 ◽

Cited By ~ 2

Author(s):

Yuanbing Wu ◽

Ania Rashidpour ◽

María Pilar Almajano ◽

Isidoro Metón

Keyword(s):

Drug Delivery ◽

Gene Therapy ◽

Nucleic Acid ◽

Gene Delivery ◽

Gonadal Development ◽

Endosomal Escape ◽

Cell Uptake ◽

Nucleic Acid Delivery ◽

Farmed Fish ◽

Therapy Studies

Chitosan is increasingly used for safe nucleic acid delivery in gene therapy studies, due to well-known properties such as bioadhesion, low toxicity, biodegradability and biocompatibility. Furthermore, chitosan derivatization can be easily performed to improve the solubility and stability of chitosan–nucleic acid polyplexes, and enhance efficient target cell drug delivery, cell uptake, intracellular endosomal escape, unpacking and nuclear import of expression plasmids. As in other fields, chitosan is a promising drug delivery vector with great potential for the fish farming industry. This review highlights state-of-the-art assays using chitosan-based methodologies for delivering nucleic acids into cells, and focuses attention on recent advances in chitosan-mediated gene delivery for fish biotechnology applications. The efficiency of chitosan for gene therapy studies in fish biotechnology is discussed in fields such as fish vaccination against bacterial and viral infection, control of gonadal development and gene overexpression and silencing for overcoming metabolic limitations, such as dependence on protein-rich diets and the low glucose tolerance of farmed fish. Finally, challenges and perspectives on the future developments of chitosan-based gene delivery in fish are also discussed.

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Zn-promoted gene transfection efficiency for non-viral vectors: a mechanism study

New Journal of Chemistry ◽

10.1039/d1nj02115j ◽

2021 ◽

Author(s):

Xiao-Qi Yu ◽

Rui-Mo Zhao ◽

Yu Guo ◽

Hui-Zhen Yang ◽

Ji Zhang

Keyword(s):

Gene Therapy ◽

Nucleic Acid ◽

Gene Delivery ◽

Viral Vectors ◽

Transfection Efficiency ◽

Gene Transfection ◽

Viral Gene ◽

Mechanism Study ◽

High Affinity ◽

Gene Vectors

The development of cationic non-viral gene vectors that may overcome the obstacles in gene delivery is of great significance to gene therapy. Metallic complexes with high affinity to nucleic acid...

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Practical gene delivery system suitable for clinical gene therapy

Nature Middle East ◽

10.1038/nmiddleeast.2010.238 ◽

2010 ◽

Author(s):

Abdulhakim Mahmoud

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Delivery System ◽

Gene Delivery System ◽

Clinical Gene Therapy

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Faculty Opinions recommendation of Nucleic acid base analog FRET-pair facilitating detailed structural measurements in nucleic acid containing systems.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1158910.619167 ◽

2009 ◽

Author(s):

Walter Chazin

Keyword(s):

Nucleic Acid ◽

Acid Base ◽

Nucleic Acid Base ◽

Base Analog ◽

Fret Pair

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Faculty Opinions recommendation of Peptide nucleic acid (PNA) cell penetrating peptide (CPP) conjugates as carriers for cellular delivery of antisense oligomers.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13939980.15395086 ◽

2012 ◽

Author(s):

Daniel Appella ◽

Ethan Englund

Keyword(s):

Nucleic Acid ◽

Peptide Nucleic Acid ◽

Cell Penetrating Peptide ◽

Cellular Delivery ◽

Cell Penetrating

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Exosome as a Natural Gene Delivery Vector for Cancer Treatment

Current Cancer Drug Targets ◽

10.2174/1568009620666200924154149 ◽

2020 ◽

Vol 20 (11) ◽

pp. 821-830

Author(s):

Prasad Pofali ◽

Adrita Mondal ◽

Vaishali Londhe

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Nucleic Acids ◽

Cancer Treatment ◽

Intestinal Barrier ◽

Gene Carrier ◽

Gene Delivery Vector ◽

Delivery Vector

Background: Current gene therapy vectors such as viral, non-viral, and bacterial vectors, which are used for cancer treatment, but there are certain safety concerns and stability issues of these conventional vectors. Exosomes are the vesicles of size 40-100 nm secreted from multivesicular bodies into the extracellular environment by most of the cell types in-vivo and in-vitro. As a natural nanocarrier, exosomes are immunologically inert, biocompatible, and can cross biological barriers like the blood-brain barrier, intestinal barrier, and placental barrier. Objective: This review focusses on the role of exosome as a carrier to efficiently deliver a gene for cancer treatment and diagnosis. The methods for loading of nucleic acids onto the exosomes, advantages of exosomes as a smart intercellular shuttle for gene delivery and therapeutic applications as a gene delivery vector for siRNA, miRNA and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and also the limitations of exosomes as a gene carrier are all reviewed in this article. Methods: Mostly, electroporation and chemical transfection are used to prepare gene loaded exosomes. Results: Exosome-mediated delivery is highly promising and advantageous in comparison to the current delivery methods for systemic gene therapy. Targeted exosomes, loaded with therapeutic nucleic acids, can efficiently promote the reduction of tumor proliferation without any adverse effects. Conclusion: In the near future, exosomes can become an efficient gene carrier for delivery and a biomarker for the diagnosis and treatment of cancer.

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