Cell Penetrating Peptide Conjugated Chitosan for Enhanced Delivery of Nucleic Acid

Layek, Buddhadev; Lipp, Lindsey; Singh, Jagdish

doi:10.3390/ijms161226142

Cited by 76 publications

(60 citation statements)

References 143 publications

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“…“Transfection” is considered to be either the process of vector entry through the cell membrane [17] or the resulting expression from the transgene [18]. In this study, we distinguished these two aspects of transfection.…”

Section: Introductionmentioning

confidence: 99%

Effects of Circular DNA Length on Transfection Efficiency by Electroporation into HeLa Cells

et al. 2016

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The ability to produce extremely small and circular supercoiled vectors has opened new territory for improving non-viral gene therapy vectors. In this work, we compared transfection of supercoiled DNA vectors ranging from 383 to 4,548 bp, each encoding shRNA against GFP under control of the H1 promoter. We assessed knockdown of GFP by electroporation into HeLa cells. All of our vectors entered cells in comparable numbers when electroporated with equal moles of DNA. Despite similar cell entry, we found length-dependent differences in how efficiently the vectors knocked down GFP. As vector length increased up to 1,869 bp, GFP knockdown efficiency per mole of transfected DNA increased. From 1,869 to 4,257 bp, GFP knockdown efficiency per mole was steady, then decreased with increasing vector length. In comparing GFP knockdown with equal masses of vectors, we found that the shorter vectors transfect more efficiently per nanogram of DNA transfected. Our results rule out cell entry and DNA mass as determining factors for gene knockdown efficiency via electroporation. The length-dependent effects we have uncovered are likely explained by differences in nuclear translocation or transcription. These data add an important step towards clinical applications of non-viral vector delivery.

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Section: Introductionmentioning

confidence: 99%

Effects of Circular DNA Length on Transfection Efficiency by Electroporation into HeLa Cells

et al. 2016

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“…Chitosan is a natural polymer widely used in regenerative medicine, and has a GAGlike structure (Layek et al, 2015). Chitosan has a positively charged surface, which facilitates the attachment of cells with negatively charged groups (Wang et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of bone matrix gelatin/fibrin glue and chitosan/gelatin composite scaffolds for cartilage tissue engineering

Wang

Zhang

et al. 2016

Genet. Mol. Res.

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ABSTRACT. This study was designed to evaluate bone matrix gelatin (BMG)/fibrin glue and chitosan/gelatin composite scaffolds for cartilage tissue engineering. Chondrocytes were isolated from costal cartilage of Sprague-Dawley rats and seeded on BMG/fibrin glue or chitosan/gelatin composite scaffolds. After different in vitro culture durations, the scaffolds were subjected to hematoxylin and eosin, Masson's trichrome, and toluidine blue staining, anti-collagen II and anti-aggrecan immunohistochemistry, and scanning electronic microscopy (SEM) analysis. After 2 weeks of culture, chondrocytes were distributed evenly on the surfaces of both scaffolds. Cell numbers and the presence of extracellular matrix components were markedly increased after 8 weeks of culture, and to a greater extent on the chitosan/gelatin scaffold. The BMG/fibrin glue scaffold showed signs of degradation after 8 weeks. Immunofluorescence analysis confirmed higher levels of collagen II and aggrecan using the chitosan/gelatin scaffold. SEM revealed that the majority of cells on the surface of the BMG/fibrin glue scaffold demonstrated a round morphology, while those in the chitosan/gelatin group had a spindle-like shape, with pseudopodia. Chitosan/gelatin scaffolds appear to be superior to BMG/ fibrin glue constructs in supporting chondrocyte attachment, proliferation, and biosynthesis of cartilaginous matrix components.

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“…Furthermore, because chitosan is positively charged, it has the ability to interact with and protect negatively charged nucleic acids such as siRNA. [24,25] However, a significant challenge to chitosan research is its variability in composition and molecular weight, which makes it difficult to understand which properties are associated with which effect. [25] Chitosan transfection efficiency is sensitive to the pH of its surroundings, and as pH cannot be controlled in vivo, more research needs to be conducted to alleviate potential problems with the vector's transfection efficiency.…”

Section: Chitosanmentioning

confidence: 99%

“…On the basis of their physicochemical characteristics, CPPs can be classified as cationic, amphipathic, or hydrophobic. [24,62] Challenges for the use of CPPs in siRNA delivery involve a potential decrease in delivery efficacy when positively-charged CPPs are linked to negatively-charged siRNA. Additionally, there is concern over CPPs having cytotoxicity or inducing immunogenicity.…”

Section: Cell-penetrating Peptidesmentioning

confidence: 99%

An Overview of Various Carriers for siRNA Delivery

Marquez¹,

Madu²,

Lü³

2018

Oncomedicine

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RNA interference through the use of short interfering molecules known as short interfering RNA (siRNA) has the potential to greatly advance research in treatments for many diseases because it has the ability to silence the expression of specific genes by helping degrade target mRNA. However, challenges to siRNA delivery have made the development of safe and effective delivery systems paramount in siRNA research. Various types of delivery systems have been proposed and investigated for siRNA delivery and therapy. Although viral vectors have been established to be the most effective in delivering siRNA molecules, they also raise many concerns over biosafety, especially concerning immunogenicity. Therefore, many researchers have begun to investigate and study non-viral vectors. Non-viral vectors are studied because they are typically considered to be safer than viral vectors albeit less efficient as well. The three general non-viral vectors that have been studied for siRNA delivery are lipid-based, non-lipid organic-based, and non-lipid inorganic-based carriers. Within those general parameters of non-viral vector classification are subtypes that are each unique with their own characteristic benefits and downsides. Many of these carriers, as well as even naked siRNA, do have the potential to be modified so that siRNA delivery could be further enhanced with benefits such as greater stability and duration. Researchers still must be wary with alterations as to not interfere with siRNA function. Currently, widespread siRNA therapeutics are still out of reach, but as more advancements in siRNA research including research on their delivery mechanisms are established, the goal of integrating siRNA therapy into the treatment of a multitude of diseases becomes increasingly more of a possibility. Researchers are currently investigating how siRNA can be used to not just treat cancer but ocular and neurodegenerative diseases as well as many others. There are still many obstacles to face and overcome before siRNA therapy can be implemented into the treatment of many diseases, and more research must still be conducted concerning siRNA delivery systems. Many advancements pertaining to siRNA carriers have been made, and many more are likely on their way.

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Cell Penetrating Peptide Conjugated Chitosan for Enhanced Delivery of Nucleic Acid

Cited by 76 publications

References 143 publications

Effects of Circular DNA Length on Transfection Efficiency by Electroporation into HeLa Cells

Effects of Circular DNA Length on Transfection Efficiency by Electroporation into HeLa Cells

Evaluation of bone matrix gelatin/fibrin glue and chitosan/gelatin composite scaffolds for cartilage tissue engineering

An Overview of Various Carriers for siRNA Delivery

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