Nucleic Acid Delivery Research Articles

Cell-Penetrating Peptide-Mediated Delivery of Gene-Silencing Nucleic Acids to the Invasive Common Reed Phragmites australis via Foliar Application

As a popular tool for gene function characterization and gene therapy, RNA interference (RNAi)-based gene silencing has been increasingly explored for potential applications to control invasive species. At least two major hurdles exist when applying this approach to invasive plants: (1) the design and screening of species- and gene-specific biomacromolecules (i.e., gene-silencing agents or GSAs) made of DNA, RNA, or peptides that can suppress the expression of target genes efficiently, and (2) the delivery vehicle needed to penetrate plant cell walls and other physical barriers (e.g., leaf cuticles). In this study, we investigated the cell-penetrating peptide (CPP)-mediated delivery of multiple types of GSAs (e.g., double-stranded RNA (dsRNA), artificial microRNA (amiRNA), and antisense oligonucleotide (ASO)) to knock down a putative phytoene desaturase (PDS) gene in the invasive common reed (Phragmites australis spp. australis). Both microscopic and quantitative gene expression evidence demonstrated the CPP-mediated internalization of GSA cargos and transient suppression of PDS expression in both treated and systemic leaves up to 7 days post foliar application. Although various GSA combinations and application rates and frequencies were tested, we observed limitations, including low gene-silencing efficiency and a lack of physiological trait alteration, likely owing to low CPP payload capacity and the incomplete characterization of the PDS-coding genes (e.g., the recent discovery of two PDS paralogs) in P. australis. Our work lays a foundation to support further research toward the development of convenient, cost-effective, field-deployable, and environmentally benign gene-silencing technologies for invasive P. australis management.

Microfluidics-Assembled Nanovesicles for Nucleic Acid Delivery.

ConspectusMicrofluidic technologies have become a highly effective platform for the precise and reproducible production of nanovesicles used in drug and nucleic acid delivery. One of their key advantages lies in the one-step assembly of multidrug delivery nanovesicles, which improves batch-to-batch reproducibility by minimizing the intermediate steps typically required in conventional methods. These steps often involve complex hydrophobic and electrostatic interactions, leading to variability in the nanovesicle composition and performance. Microfluidic systems streamline the encapsulation of diverse therapeutic agents, including hydrophilic nucleic acids, proteins, and both hydrophobic and hydrophilic small molecules, within a single chip, ensuring a more consistent production process. This capability enables the codelivery of multiple drugs targeting different disease pathways, which is particularly valuable in reducing the risk of drug resistance.Despite the promise of nanovesicles for nucleic acid delivery, their clinical translation has been hindered by safety concerns, particularly cytotoxicity, which has overshadowed efforts to improve in vivo stability and delivery efficiency. Positively charged nanovesicles, commonly used to encapsulate negatively charged nucleic acids, tend to exhibit significant cytotoxicity. To address this, charge-shifting materials that respond to pH changes or surface modifications have been proposed as promising strategies. Shifting the surface charge from positive to neutral or negative at physiological pH can reduce the cytotoxicity, enhancing the clinical feasibility of these nanovesicle-based therapies.Microfluidic platforms offer precise control over key nanovesicle properties, including particle size, rigidity, morphology, and encapsulation efficiency. Particle size is relatively easy to adjust by controlling flow rates within microfluidic channels, with higher flow rates generally producing smaller particles. However, continuous tuning of the particle rigidity remains challenging. By manipulation of the interfacial water layer between hydrophobic and amphiphilic components during nanoparticle formation, future designs may achieve greater control over rigidity, which is critical for improving cellular uptake and biodistribution. While shape tuning using microfluidic chips has not yet been fully explored in biomedical applications, advances in materials science may enable this aspect in the future, offering further customization of the nanovesicle properties.The integration of nanovesicle assembly and surface modification within a single microfluidic platform presents challenges due to the differing speeds of these processes. Nanovesicle assembly is typically rapid, whereas surface modifications, such as those involving functional biomolecules, occur more slowly and often require purification steps. Recent advances, such as rotary valve designs and single-axis camshaft mechanisms, offer precise control over flow mixing at different stages of the process, allowing for the automation of nanovesicle assembly and surface modification, thereby improving batch-to-batch reproducibility.In conclusion, microfluidic technologies represent a promising approach for the development of multifunctional nanovesicles with the potential to address key challenges in drug delivery and precision medicine. While obstacles related to cytotoxicity, scalability, and reproducibility remain, innovations in chip design, materials, and automation are paving the way for broader application in clinical settings. Future research, potentially incorporating machine learning, could further optimize the relationship between nanovesicle properties and biological outcomes, advancing the use of microfluidic technologies for therapeutic delivery.

Improving lipid nanoparticles delivery efficiency of macrophage cells by using immunomodulatory small molecules.

Nucleic acid drug delivery remains a key challenge in the development of nucleic acid therapy. How to improve the efficiency of nucleic acid delivery is still an important strategy for Lipofectamine 3000 and LNP development. Here, we screened 248 inhibitors or agonists related to immune modulation and identified three small molecules (BP-1-102, SCH58261, and Bropirimine) that could enhance the transfection efficiency to 2-fold to 5-fold of Lipofectamine 3000 and LNP, all of which are currently approved for clinical use in the treatment of the most common malignant tumors. In addition, we used high-throughput RNA sequencing technology to analyze the mechanisms and found that they were mainly associated with the receptor-mediated endocytosis pathway. Our findings have yielded novel insights that can contribute to the advancement of nucleic acid drugs, enhancing both their efficacy and precision in targeting cancer therapy.

Turning waste into wealth: Enzyme-activated DNA sensor based on reactant recycle for spatially selective imaging microRNA toward target cells.

Amplified imaging of microRNA (miRNA) in cancer cells is essential for understanding of the underlying pathological process. Synthetic catalytic DNA circuits represent pivotal tools for miRNA imaging. However, most existing catalytic DNA circuits can not achieve the reactant recycling operation in cells and in vivo. Additionally, specificity miRNA imaging in tumor site also is a challenge. Herein, inspired by "turning waste into wealth", we report a DNA sensor for imaging of miRNA in target cells based on apurinic/apyrimidinic endonuclease 1 (APE1)-activated reactant recycling catalytic DNA circuit. The sensing function of the DNA sensor is suppressed firstly through engineering an apurinic/apyrimidinic (AP) site. In the presence of APE1, the AP site undergoes hydrolysis, thereby activating sensing activity and triggering the strand displacement reaction (SDR) under miRNA assistance. In this catalytic DNA circuit, the next reaction cycle can be initiated when the duplex strand waste products are reverted into active components, which allows it to be performed continuously just consuming fuel DNA without depleting the reactant. Noteworthy, the liposome plays important role in overcome biological barriers for nucleic acid delivery. The amplified miRNA imaging strategy is carried out in vivo by this DNA sensor with reducing off-tumor signal under assistance with APE1, and enhances tumor-to-normal tissue contrast compared with traditional catalytic DNA circuit. Firstly, APE1-activated reactant recycling catalytic DNA circuit is developed. Secondly, miRNA image in cell and in animal is achieved with high spatial selectivity. Thirdly, the signal-to-background ratio for imaging is improved in vitro and in vivo. Lastly, our strategy provides an automatic yet versatile approach for the development of more efficient and selective DNA circuits capable of differentiating miRNA in cancer cells from those in normal cells, promising to be valuable in cancer diagnosis.

Effects of Electroporation Timing and Cumulus Cell Attachment on InVitro Development and Genome Editing of Porcine Embryos.

Pig genome editing using the oviductal nucleic acid delivery (GONAD) method with electroporation would allow the efficient obtention of genetically modified pigs. However, oocytes and zygotes at various stages after ovulation must be targeted, and cumulus cell attachment and mosaic mutations are major obstacles. Therefore, we investigated whether two parameters (electroporation timing and the cumulus cell attachment) influence the effectiveness of multiplex genome editing by electroporation in porcine oocytes or zygotes. Three gRNAs targeting either GGTA1, CMAH or B4GALNT2 were introduced individually into oocytes and zygotes with and without cumulus cells at three different time points, 0 h before invitro fertilisation (IVF) and 5 h and 10 h after IVF initiation. The introduction of gRNAs into oocytes and zygotes did not significantly affect the rates of blastocyst formation and total mutation of the resulting blastocysts irrespective of cumulus cell attachment and electroporation timing. In conclusion, the electroporation timing and the cumulus cell attachment did not interfere with the efficient delivery of the CRISPR/Cas9 system to the oocytes/zygotes, indicating that porcine genome editing in the oviduct using GONAD method may be possible.

Current landscape of metal–organic framework-mediated nucleic acid delivery and therapeutics

Current landscape of metal–organic framework-mediated nucleic acid delivery and therapeutics

RETRACTION: Delivery of Nucleic Acids Through the Controlled Disassembly of Multifunctional Nanocomplexes

RETRACTION: Delivery of Nucleic Acids Through the Controlled Disassembly of Multifunctional Nanocomplexes

Applications of liposomes and lipid nanoparticles in cancer therapy: current advances and prospects.

Liposomes and lipid nanoparticles are common lipid-based drug delivery systems and play important roles in cancer treatment and vaccine manufacture. Although significant progress has been made with these lipid-based nanocarriers in recent years, efficient clinical translation of active targeted liposomal nanocarriers remains extremely challenging. In this review, we focus on targeted liposomes, stimuli-responsive strategy and combined therapy in cancer treatment. We also summarize advances of liposome and lipid nanoparticle applications in nucleic acid delivery and tumor vaccination. In addition, we discuss limitations and challenges in the clinical translation of these lipid nanomaterials and make recommendations for the future research in cancer therapy.

Smart Stimuli-responsive Nanogels: A Potential Tool for Targeted Drug Delivery.

Nanogels (NGs) are presently the focus of extensive research because of their special qualities, including minimal particle size, excellent encapsulating efficacy, and minimizing the breakdown of active compounds. As a result, NGs are great candidates for drug delivery systems. Cross-linked nanoparticles (NPs) called stimulus-responsive NGs are comprised of synthetic, natural, or a combination of natural and synthetic polymers. These NPs can swell in response to large amounts of solvent, but their structural makeup prevents them from dissolving. Furthermore, in response to (i) physical stimuli like temperatures, ion strength, and magnetized or electrical fields; (ii) chemical stimuli like the pH level, molecules, or ions; (iii) biological stimuli like the enzymatic substrate or affinity ligand, they transform into a hard particle (collapsed form) from a polymer solution (swell form). Over the past decade, there has been a major advancement in the creation of "smart" NGs in applications related to therapeutics and diagnosis, involving nucleic acid and intracellular drug delivery, photodynamic/photothermal treatment, biological imaging, and its detection. The nanogels reviewed in this article rely only on temperatures, pH, light, magnetic fields, and combinations of those variables. Developing a targeted delivery vehicle will greatly benefit from the presented information, especially when used for Core-shell multi-sensitive photo-sensitive nanogels.

Cationic‐Hydrophilic Di‐Block Copolymers: Surface‐Shielded Vectors for Gene Delivery

ABSTRACTTo tackle the main challenges in gene therapy, synthetic polycationic vectors are developed for nucleic acid (NA) delivery. In this work, vectors based on di‐block copolymers consisting of a methacrylate‐type cationic block and an electroneutral hydrophilic block are designed and synthesized to condense NA into compact particles shielded with a bioinert hydrophilic polymer layer. The results confirm that the cationic vector blocks, containing permanently charged trimethylammonium (TMA) or tributylphosphonium (TBP) groups and 0–20 mol% hydrophobic butyl groups, are able to efficiently complex DNA to form 100 to 200 nm spheroids, while the hydrophilic blocks based on 2‐methacryloyloxyethylphosphorylcholine or N‐(2‐hydroxypropyl) methacrylamide coat the nanoparticle surfaces with an electroneutral polymer layer. Additionally, pH‐responsive hydrazone bonds are incorporated between the TMA and TBP groups and the main polymer backbone of the polycationic block to ensure hydrolysis of the hydrazone bond. Theoretically, this should be accompanied by DNA release from the complex in a mildly acidic environment inside the cells while maintaining its stability under neutral blood conditions. In summary, this model study provides insight into the process of complexation/decomplexation of NA by pH‐responsive di‐block polycationic vectors. The results may contribute to the development of safe and efficient delivery systems for gene therapy applications.

Mannose-modified exosomes loaded with MiR-23b-3p target alveolar macrophages to alleviate acute lung injury in Sepsis.

The anti-inflammatory role of miR-23b-3p (miR-23b) is known in autoimmune diseases like multiple sclerosis, systemic lupus erythematosus, and rheumatoid arthritis. However, its role in sepsis-related acute lung injury (ALI) and its effect on macrophages in ALI remain unexplored. This investigation aimed to evaluate miR-23b's therapeutic potential in macrophages in the context of ALI. The study found reduced miR-23b expression in macrophages within ALI tissue. Intratracheal delivery of miR-23b mimics alleviated ALI by partially inhibiting M1 macrophage activation through the Lpar1-NF-κB pathway. Effective delivery systems are crucial for prolonging miR-23b activity in the lungs, reducing dosage, and minimizing side effects by specifically targeting macrophages. However, current vector systems for nucleic acid delivery, including viral, lipid-based, polymer-based, and peptide-based vectors, face limitations due to eliciting immune responses. Exosomes have garnered significant attention as a leading gene delivery system due to the safety, effectivity and low immunogenicity. We further isolated exosomes from bone marrow-derived mesenchymal stem cells, modified exosomes with mannosylated ligands to enhance the targeted delivery of miR-23b to macrophage. This approach represents a promising novel therapeutic strategy for treating sepsis-induced ALI.

DNA-loaded targeted nanoparticles as a safe platform to produce exogenous proteins in tumor B cells

IntroductionThe functionalization of nanoparticles (NPs) with an antiCD19 targeting mechanism represents a promising approach for the selective delivery of drugs and nucleic acids into normal and tumor B cells. This strategy has the advantage of minimizing off-target effects by restricting gene delivery to the desired cell population. However, the nanoplatform must guarantee both the local production of the protein and the safety of the treatment to allow an effective therapy with reduced systemic toxicity.MethodsIn order to ensure a selective delivery of nucleic acids, we developed poly(lactic-co-glycolic acid) (PLGA)-poly(vinyl alcohol) (PVA) NPs loaded with an Enhanced Green Fluorescent Protein (EGFP)-coding plasmid and covalently coated with antiCD19 recombinant antibody as a targeting mechanism. To assess the functionality of the NPs, physicochemical characterization, safety tests, and transfection assay were employed to evaluate the NPs’ behavior in vitro and in vivo, in a human/zebrafish lymphoma xenograft model.ResultsThe results demonstrated that the PLGA-PVA nanoplatform was capable of efficiently encapsulating and releasing the payload. These nanostructures demonstrated a favorable safety profile, as evidenced by the absence of significant cell cytotoxicity, coagulation activation, complement system activation, and the slight activation of endothelial cells and leukocytes. The targeting mechanism facilitated the interaction of NPs with target cells, thereby enhancing their internalization and subsequent exogenous plasmid DNA (pDNA) translation and protein expression. In the human/zebrafish lymphoma xenograft model, no evidence of toxicity was observed, and targeted NPs demonstrated the capacity to enhance exogenous pDNA expression.ConclusionOur findings provide a rationale for the use of targeted NPs as a DNA delivery system for the local expression of therapeutic proteins.

Enhancing non-viral gene delivery to human T cells through tuning nanoparticles physicochemical features, modulation cellular physiology, and refining transfection strategies.

Genetically engineered immune cells hold great promise for treating immune-related diseases, but their development is hindered by technical challenges, primarily related to nucleic acid delivery. Polyethylenimine (PEI) is a cost-effective transfection agent, yet it requires significant optimization for effective T cell transfection. In this study, we comprehensively fine-tuned the characteristics of PEI/DNA nanoparticles, culture conditions, cellular physiology, and transfection protocols to enhance gene delivery into T cells. Gel retardation and dynamic light scattering (DLS) analyses confirmed that PEI effectively bound to DNA, forming size- and charge-adjustable particles based on the N/P ratio, which remained stable in RPMI 1640 medium for 3 days at 25°C. At an N/P ratio of 8.0, these nanoparticles achieved an optimal transfection rate, which improved further with adjustments in DNA dosage and complex volume. Additionally, increasing the cell seeding density and adding complete media shortly after transfection significantly boosted PEI-mediated gene delivery. Notably, reversing the transfection in vials resulted in a 20-fold increase in cellular uptake and transfection efficiency compared to the conventional direct transfection method in culture plates. Finally, modifying cellular physiology with hypotonic extracellular media at pH 9.0 dramatically enhanced transfection rates while maintaining minimal cytotoxicity. These findings could reduce the cost and complexity of preparing engineered T cells, potentially accelerating the development of immune cell therapies for human diseases.

Multiplexed siRNA Immunoassay Unveils Spatial and Quantitative Dimensions of siRNA Function, Abundance, and Localization In Vitro and In Vivo.

Small interfering RNAs (siRNAs) have been successfully used as therapeutics to silence disease-causing genes when conjugated to ligands or formulated in lipid nanoparticles to target relevant cell types for efficacy while sparing other cells for safety. To support the development of new methods for delivery of siRNA therapeutics, we developed and characterized a panel of antibodies generated against chemically modified nucleotides used in therapeutic siRNA molecules, identifying a monoclonal antibody that detects a broad range of siRNA representing distinct sequences and modification patterns. By integrating this anti-siRNA antibody with additional reagents, we created a multiplex siRNA immunoassay that simultaneously quantifies siRNA uptake, trafficking, and silencing activity. Using immunohistochemistry (IHC), we applied our method on tissues from mice treated with unconjugated, GalNAc-conjugated, or cholesterol-conjugated siRNAs and quantitatively assessed the biodistribution and activity of siRNAs in various organs. In addition, we used high-content imaging (HCI) and applied our multiplex siRNA immunoassay in tissue culture to enable simultaneous quantification of siRNA uptake, activity, and intracellular colocalization with endosome markers. These methods provide a robust platform for testing nucleic acid delivery methods in vitro and in vivo, allowing precise analysis and visualization of the pharmacokinetics and pharmacodynamics of siRNA therapeutics with cellular and subcellular resolution.

Chemistry, manufacturing and controls strategies for using novel excipients in lipid nanoparticles.

Lipid nanoparticles (LNPs) for nucleic acid delivery often use novel lipids as functional excipients to modulate the biodistribution, pharmacokinetics, pharmacodynamics and efficacy of the nucleic acid. Novel excipients used in pharmaceutical products are subject to heightened regulatory scrutiny and often require data packages comparable to an active pharmaceutical ingredient. Although these regulatory requirements may help to ensure patient safety they also create economic and procedural barriers that can disincentivize innovation and delay clinical investigation. Despite the unique structural and functional role of lipid excipients in LNPs, there is limited specific global regulatory guidance, which adds uncertainty and risk to the development of LNPs. In this Perspective we provide an industry view on the chemistry, manufacturing and controls challenges that pharmaceutical companies face in the use of novel lipid excipients at each stage of development, and propose consensus recommendations on how to streamline and clarify development and regulatory expectations.

Jet Injection of Naked mRNA Encoding the RBD of the SARS-CoV-2 Spike Protein Induces a High Level of a Specific Immune Response in Mice.

Background: Although mRNA vaccines encapsulated in lipid nanoparticles (LNPs) have demonstrated a safety profile with minimal serious adverse events in clinical trials, there is opportunity to further reduce mRNA reactogenicity. The development of naked mRNA vaccines could improve vaccine tolerability. Naked nucleic acid delivery using the jet injection method may be a solution. Methods: In the first part of the study, the optimal conditions providing low traumatization and high expression of the model mRNA-GFP molecule in the tissues of laboratory animals were determined. Then, we used the selected protocol to immunize BALB/c mice with mRNA-RBD encoding the SARS-CoV-2 receptor-binding domain (RBD). It was demonstrated that mice vaccinated with naked mRNA-RBD developed a high level of specific antibodies with virus-neutralizing activity. The vaccine also induced a strong RBD-specific T-cell response and reduced the viral load in the lungs of the animals after infection with the SARS-CoV-2 virus. The level of immune response in mice immunized with mRNA-RBD using a spring-loaded jet injector was comparable to that in animals immunized with mRNA-RBD encapsulated in LNPs. Results: In this study, the efficacy of an inexpensive, simple, and safe method of mRNA delivery using a spring-loaded jet injector was evaluated and validated. Conclusions: Our findings suggest that the jet injection method may be a possible alternative to LNPs for delivering mRNA vaccines against SARS-CoV-2 infection.

Non-Viral Delivery Systems to Transport Nucleic Acids for Inherited Retinal Disorders.

Inherited retinal disorders (IRDs) represent a group of challenging genetic conditions that often lead to severe visual impairment or blindness. The complexity of these disorders, arising from their diverse genetic causes and the unique structural and functional aspects of retinal cells, has made developing effective treatments particularly challenging. Recent advancements in gene therapy, especially non-viral nucleic acid delivery systems like liposomes, solid lipid nanoparticles, dendrimers, and polymersomes, offer promising solutions. These systems provide advantages over viral vectors, including reduced immunogenicity and enhanced targeting capabilities. This review delves into introduction of common IRDs such as Leber congenital amaurosis, retinitis pigmentosa, Usher syndrome, macular dystrophies, and choroideremia and critically assesses current treatments including neuroprotective agents, cellular therapy, and gene therapy along with their limitations. The focus is on the emerging role of non-viral delivery systems, which promise to address the current limitations of specificity, untoward effects, and immunogenicity in existing gene therapies. Additionally, this review covers recent clinical trial developments in gene therapy for retinal disorders.

Direct cytosol delivery of mRNA by micron-sized co-assembly with designer oligopeptides.

Inefficient endosomal escape has been regarded as the main bottleneck for intracellular nucleic acid delivery. While most research efforts have been spent on designing various nano-sized particles, we took a different path here, investigating micron-sized carriers for direct cytosol entry. Using the spontaneous co-assembly of mRNA and the designer 27 amino acid oligopeptide named pepMAX2, micron-sized co-assemblies were obtained with various sizes by altering the concentration of NaCl salt and time for pre-incubation. Surprisingly, transfection was much more effective using micron-sized than nano-sized co-assemblies, and the efficiency surpasses that of a widely used lipid-based commercial reagent. The study was complemented by computational simulations, inhibitor studies and live-cell confocal imaging to shed light on the role of electrostatic interaction on assembly and the mechanism of uptake and intracellular trafficking. These micron-sized co-assemblies directly enter the cytosol and then release mRNA, bypassing conventional pathways and thus avoiding the lysosomal degradation. This simple approach involving short oligopeptides and salt addition to create optimal micron-sized co-assembly with mRNA should open new avenues to overcome endosomal barriers for intracellular delivery of nucleic acid therapeutics.

Photochemical Stabilization of Self-Assembled Spherical Nucleic Acids.

Oligonucleotide therapeutics, including antisense oligonucleotides and small interfering RNA, offer promising avenues for modulating the expression of disease-associated proteins. However, challenges such as nuclease degradation, poor cellular uptake, and unspecific targeting hinder their application. To overcome these obstacles, spherical nucleic acids have emerged as versatile tools for nucleic acid delivery in biomedical applications. Our laboratory has introduced sequence-defined DNA amphiphiles which self-assemble in aqueous solutions. Despite their advantages, self-assembled SNAs can be inherently fragile due to their reliance on non-covalent interactions and fall apart in biologically relevant conditions, specifically by interaction with serum proteins. Herein, this challenge is addressed by introducing two methods of covalent crosslinking of SNAs via UV irradiation. Thymine photodimerization or disulfide crosslinking at the micellar interface enhance SNA stability against human serum albumin binding. This enhanced stability, particularly for disulfide crosslinked SNAs, leads to increased cellular uptake. Furthermore, this crosslinking results in sustained activity and accessibility for release of the therapeutic nucleic acid, along with improvement in unaided gene silencing. The findings demonstrate the efficient stabilization of SNAs through UV crosslinking, influencing their cellular uptake, therapeutic release, and ultimately, gene silencing activity. These studies offer promising avenues for further optimization and exploration of pre-clinical, in vivo studies.

Cationic Cyclodextrin-Based Carriers for Drug and Nucleic Acid Delivery.

Cyclodextrins can serve as carriers for various payloads, utilizing their capacity to form unique host-guest inclusion complexes within their cavity and their versatile surface functionalization. Recently, cationic cyclodextrins have gained considerable attention, as they can improve drug permeability across negatively charged cell membranes and efficiently condense negatively charged nucleic acid due to electrostatic interactions. This review focuses on state-of-the-art and recent advances in the construction of cationic cyclodextrin-based delivery systems. First, we identified different cationic moieties that are commonly employed in the design of cyclodextrins with enhanced complexation ability. Subsequently, a wide range of cationic cyclodextrin-based drug delivery systems were analyzed with emphasis on chemistry, drug release profiles, and therapeutic outcomes. The evaluation of the delivery platforms was also based on the four major types of drugs, such as anticancer, anti-inflammatory, antibacterial, and antidiabetic agents. The delivery systems for nucleic acids were also summarized while focusing on their condensation ability, transfection efficiency, and biocompatibility in comparison to commercially available vectors such as PEI 25 kDa and lipofectamine 2000. Furthermore, we highlighted the potential of cationic cyclodextrins in constructing multimodal delivery systems for the simultaneous encapsulation of both drugs and nucleic acids. Finally, the challenges and limitations associated with cationic cyclodextrin setups were discussed.