One-step encapsulation of siRNA between lipid-layers of multi-layer polycation liposomes by lipoplex freeze-thawing

Koide, Hiroyuki; Okamoto, Ai; Tsuchida, Hiroki; Ando, Hidenori; Ariizumi, Saki; Kiyokawa, Chiaki; Hashimoto, Masahiro; Asai, Tomohiro; Dewa, Takehisa; Oku, Naoto

doi:10.1016/j.jconrel.2016.01.032

Cited by 23 publications

(25 citation statements)

References 31 publications

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“…The first two applied methods are gel electrophoresis and fluorescence‐based assay (RiboGreen ® ). Indeed, these methods are commonly used and have been largely described for the quantification of siRNA . The third method is CE, which is generally more described for the determination and the separation of different oligonucleotides, mainly DNA [, ] rather than for the evaluation of the liposomes siRNA complexation efficiency.…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, efficient and successful siRNA delivery into target cells is the most challenging step for clinical application. Indeed, naked siRNA is susceptible to rapid degradation and its high molecular weight and negatively charged nature prevent its association with cell membranes . In order to overcome these barriers, siRNA needs to be incorporated in an adequate vector.…”

Section: Introductionmentioning

confidence: 99%

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Capillary electrophoresis method to determine siRNA complexation with cationic liposomes

et al. 2016

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Small interfering RNA (siRNA) inducing gene silencing has great potential to treat many human diseases. To ensure effective siRNA delivery, it must be complexed with an appropriate vector, generally nanoparticles. The nanoparticulate complex requires an optimal physiochemical characterization and the complexation efficiency has to be precisely determined. The methods usually used to measure complexation in gel electrophoresis and RiboGreen fluorescence-based assay. However, those approaches are not automated and present some drawbacks such as the low throughput and the use of carcinogenic reagents. The aim of this study is to develop a new simple and fast method to accurately quantify the complexation efficiency. In this study, capillary electrophoresis (CE) was used to determine the siRNA complexation with cationic liposomes. The short-end injection mode applied enabled siRNA detection in less than 5 min. Moreover, the CE technique offers many advantages compared with the other classical methods. It is automated, does not require sample preparation and expensive reagents. Moreover, no mutagenic risk is associated with the CE approach since no carcinogenic product is used. Finally, this methodology can also be extended for the characterization of other types of nanoparticles encapsulating siRNA, such as cationic polymeric nanoparticles.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis method to determine siRNA complexation with cationic liposomes

et al. 2016

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“…Serum stability assay was performed to determine the dendrimer mediated protection of siPLK1 in presence of serum (Koide et al, ). For this purpose, siPLK1 dendriplexes with CPD 3, CPD 4, PAMAM 3, and PAMAM 4 at 3:1 N/P ratio were prepared, mixed with equal volume of DMEM supplemented with 10% of FBS and incubated at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Dendrimer mediated targeting of siRNA against polo‐like kinase for the treatment of triple negative breast cancer

Jain

Mahira

Majoral

et al. 2019

J Biomedical Materials Res

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Irresponsiveness of triple negative breast cancer (TNBC) toward conventional therapies has drawn attention toward siRNA therapeutics. In gene delivery, dendrimers are gaining significant attention due to their characteristic features and polo‐like kinase (PLK1) is reported as a potential target for TNBC. In this work, phosphorus and polyamidoamine dendrimer (generation 3 and 4 of each type) are explored to address delivery challenges of PLK1 siRNA (siPLK1). Dendriplexes were formed and complexation was found at 3:1 N/P ratio for all dendrimers by gel electrophoresis. Complexation was also supported by zeta potential, circular dichroism and intercalation assay. Dendriplexes were found to be stable in presence of ribonuclease and serum. Dendriplexes resulted in enhanced cell uptake of siPLK1 compared to siPLK1 solution in MDA‐MB‐231 and MCF‐7 cells. Dendriplexes caused increased cell arrest in sub‐G1 phase compared to solution. These observations suggested phosphorus and polyamidoamine dendrimers as potential carriers for siPLK1 delivery to treat TNBC.

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“…These findings were attributed to the proton sponge effect of TEPA-PCL [38]. Liposomes containing the polycation dicetyl phosphate-diethylenetriamine (DCP-DETA) have also been proposed by Koide et al to deliver siRNA [39]. They reported that the DOPE/chol/DCP-DETA CLs were firstly compexed with siRNA and then freeze-thawed.…”

Section: Lipid Vesiclesmentioning

confidence: 99%

“…Following freeze-thawing of a single-layer CLs/siRNA complex, a “packed multi-layer structure” is formed, suggesting that siRNA was effectively encapsulated between the lipid layers. Interestingly, no degradation of encapsulated siRNA in freeze-thawed lipoplex was found in bovine serum for 72 h, while, in the same experimental conditions, 90% of siRNA in the conventional lipoplex was degraded [39]. …”

Section: Lipid Vesiclesmentioning

confidence: 99%

Lipid Nanovectors to Deliver RNA Oligonucleotides in Cancer

et al. 2016

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The growing knowledge on the mechanisms of gene silencing and gene regulation by non-coding RNAs (ncRNA), mainly small interfering RNA (siRNA) and microRNA (miRNA), is providing a significant boost to the development of new therapeutic strategies for the treatment of cancer. However, the design of RNA-based therapeutics is hampered by biopharmaceutical issues, thus requiring the use of suitable delivery strategies. In this regards, lipid nanovectors have been successfully investigated to deliver RNA in different forms of cancer. Compared to other biomaterials, lipids offer advantages such as biocompatibility, biodegradability, easy production, low cost, limited toxicity and immunogenicity. The possibility to formulate these materials in the form of nanovectors allows overcoming biopharmaceutical issues associated to the therapeutic use of RNA, with the possibility to target tumors. This review takes stock of the main lipid nanovectors proposed to deliver ncRNA. For each considered delivery strategy, the rational design and the most meaningful in vitro and in vivo results are reported and discussed.

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One-step encapsulation of siRNA between lipid-layers of multi-layer polycation liposomes by lipoplex freeze-thawing

Cited by 23 publications

References 31 publications

Capillary electrophoresis method to determine siRNA complexation with cationic liposomes

Capillary electrophoresis method to determine siRNA complexation with cationic liposomes

Dendrimer mediated targeting of siRNA against polo‐like kinase for the treatment of triple negative breast cancer

Lipid Nanovectors to Deliver RNA Oligonucleotides in Cancer

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