scholarly journals Effective carrier-free gene-silencing activity of cholesterol-modified siRNAs

RSC Advances ◽  
2018 ◽  
Vol 8 (41) ◽  
pp. 22963-22966 ◽  
Author(s):  
Lidya Salim ◽  
Chris McKim ◽  
Jean-Paul Desaulniers
Keyword(s):  
Gene Silencing ◽  
Great Promise ◽  
Chemical Modifications ◽  
Pharmacokinetic Properties ◽  
Carrier Free ◽  
Free Gene ◽  
Effective Carrier

The use of short interfering RNAs (siRNAs) as therapeutics holds great promise, but chemical modifications must first be employed to improve their pharmacokinetic properties.

Cationic siRNAs Provide Carrier-Free Gene Silencing in Animal Cells

2009 ◽  
Vol 131 (49) ◽  
pp. 17730-17731 ◽  
Author(s):  
Marc Nothisen ◽  
Mitsuharu Kotera ◽  
Emilie Voirin ◽  
Jean-Serge Remy ◽  
Jean-Paul Behr
Keyword(s):  
Gene Silencing ◽  
Animal Cells ◽  
Carrier Free ◽  
Free Gene

Structure Tuning of Cationic Oligospermine–siRNA Conjugates for Carrier-Free Gene Silencing

2016 ◽  
Vol 13 (8) ◽  
pp. 2718-2728 ◽  
Author(s):  
Marc Nothisen ◽  
Jérémy Bagilet ◽  
Jean-Paul Behr ◽  
Jean-Serge Remy ◽  
Mitsuharu Kotera
Keyword(s):  
Gene Silencing ◽  
Carrier Free ◽  
Free Gene

scholarly journals Carrier-free Gene Silencing by Amphiphilic Nucleic Acid Conjugates in Differentiated Intestinal Cells

2016 ◽  
Vol 5 ◽  
pp. e364 ◽  
Author(s):  
Elena Moroz ◽  
Soo Hyeon Lee ◽  
Ken Yamada ◽  
François Halloy ◽  
Saúl Martínez-Montero ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Gene Silencing ◽  
Intestinal Cells ◽  
Carrier Free ◽  
Free Gene

scholarly journals Lipophilic siRNA targets albumin in situ and promotes bioavailability, tumor penetration, and carrier-free gene silencing

2017 ◽  
Vol 114 (32) ◽  
pp. E6490-E6497 ◽  
Author(s):  
Samantha M. Sarett ◽  
Thomas A. Werfel ◽  
Linus Lee ◽  
Meredith A. Jackson ◽  
Kameron V. Kilchrist ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Xenograft Model ◽  
Fold Increase ◽  
Cell Uptake ◽  
Orthotopic Model ◽  
Pharmacokinetic Properties ◽  
Carrier Free ◽  
Tumor Accumulation

Clinical translation of therapies based on small interfering RNA (siRNA) is hampered by siRNA's comprehensively poor pharmacokinetic properties, which necessitate molecule modifications and complex delivery strategies. We sought an alternative approach to commonly used nanoparticle carriers by leveraging the long-lived endogenous serum protein albumin as an siRNA carrier. We synthesized siRNA conjugated to a diacyl lipid moiety (siRNA-L2), which rapidly binds albumin in situ. siRNA-L2, in comparison with unmodified siRNA, exhibited a 5.7-fold increase in circulation half-life, an 8.6-fold increase in bioavailability, and reduced renal accumulation. Benchmarked against leading commercial siRNA nanocarrier in vivo jetPEI, siRNA-L2 achieved 19-fold greater tumor accumulation and 46-fold increase in per-tumor-cell uptake in a mouse orthotopic model of human triple-negative breast cancer. siRNA-L2 penetrated tumor tissue rapidly and homogeneously; 30 min after i.v. injection, siRNA-L2 achieved uptake in 99% of tumor cells, compared with 60% for jetPEI. Remarkably, siRNA-L2 achieved a tumor:liver accumulation ratio >40:1 vs. <3:1 for jetPEI. The improved pharmacokinetic properties of siRNA-L2 facilitated significant tumor gene silencing for 7 d after two i.v. doses. Proof-of-concept was extended to a patient-derived xenograft model, in which jetPEI tumor accumulation was reduced fourfold relative to the same formulation in the orthotopic model. The siRNA-L2 tumor accumulation diminished only twofold, suggesting that the superior tumor distribution of the conjugate over nanoparticles will be accentuated in clinical situations. These data reveal the immense promise of in situ albumin targeting for development of translational, carrier-free RNAi-based cancer therapies.

RNAi gene silencing using cerasome as a viral-size siRNA-carrier free from fusion and cross-linking

2007 ◽  
Vol 17 (14) ◽  
pp. 3935-3938 ◽  
Author(s):  
Kazuki Matsui ◽  
Yoshihiro Sasaki ◽  
Takayoshi Komatsu ◽  
Masaru Mukai ◽  
Jun-ichi Kikuchi ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Cross Linking ◽  
Carrier Free

scholarly journals In vitro and in vivo properties of therapeutic oligonucleotides containing non-chiral 3′ and 5′ thiophosphate linkages

2019 ◽  
Vol 48 (1) ◽  
pp. 63-74 ◽  
Author(s):  
Jörg Duschmalé ◽  
Henrik Frydenlund Hansen ◽  
Martina Duschmalé ◽  
Erich Koller ◽  
Nanna Albaek ◽  
...  
Keyword(s):  
Antisense Oligonucleotides ◽  
Rnase H ◽  
Modified Oligonucleotides ◽  
In Vitro Experiments ◽  
Chemical Modifications ◽  
Pharmacokinetic Properties ◽  
Target Binding ◽  
The Individual

Abstract The introduction of non-bridging phosphorothioate (PS) linkages in oligonucleotides has been instrumental for the development of RNA therapeutics and antisense oligonucleotides. This modification offers significantly increased metabolic stability as well as improved pharmacokinetic properties. However, due to the chiral nature of the phosphorothioate, every PS group doubles the amount of possible stereoisomers. Thus PS oligonucleotides are generally obtained as an inseparable mixture of a multitude of diastereoisomeric compounds. Herein, we describe the introduction of non-chiral 3′ thiophosphate linkages into antisense oligonucleotides and report their in vitro as well as in vivo activity. The obtained results are carefully investigated for the individual parameters contributing to antisense activity of 3′ and 5′ thiophosphate modified oligonucleotides (target binding, RNase H recruitment, nuclease stability). We conclude that nuclease stability is the major challenge for this approach. These results highlight the importance of selecting meaningful in vitro experiments particularly when examining hitherto unexplored chemical modifications.

scholarly journals siRNA-Loaded Hydroxyapatite Nanoparticles for KRAS Gene Silencing in Anti-Pancreatic Cancer Therapy

Pharmaceutics ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1428
Author(s):  
Dandan Luo ◽  
Xiaochun Xu ◽  
M. Zubair Iqbal ◽  
Qingwei Zhao ◽  
Ruibo Zhao ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Therapy ◽  
Gene Silencing ◽  
Targeted Delivery ◽  
High Efficiency ◽  
Great Promise ◽  
Kras Gene ◽  
Promising Alternative ◽  
Pancreatic Cancer Therapy

Pancreatic carcinoma (PC) is greatly induced by the KRAS gene mutation, but effective targeted delivery for gene therapy has not existed. Small interfering ribonucleic acid (siRNA) serves as an advanced therapeutic modality and holds great promise for cancer treatment. However, the development of a non-toxic and high-efficiency carrier system to accurately deliver siRNA into cells for siRNA-targeted gene silencing is still a prodigious challenge. Herein, polyethylenimine (PEI)-modified hydroxyapatite (HAp) nanoparticles (HAp-PEI) were fabricated. The siRNA of the KRAS gene (siKras) was loaded onto the surface of HAp-PEI via electrostatic interaction between siRNA and PEI to design the functionalized HAp-PEI nanoparticle (HAp-PEI/siKras). The HAp-PEI/siKras was internalized into the human PC cells PANC-1 to achieve the maximum transfection efficiency for active tumor targeting. HAp-PEI/siKras effectively knocked down the expression of the KRAS gene and downregulated the expression of the Kras protein in vitro. Furthermore, the treatment with HAp-PEI/siKras resulted in greater anti-PC cells’ (PANC-1, BXPC-3, and CFPAC-1) efficacy in vitro. Additionally, the HAp-PEI exhibited no obvious in vitro cytotoxicity in normal pancreatic HPDE6-C7 cells. These findings provided a promising alternative for the therapeutic route of siRNA-targeted gene engineering for anti-pancreatic cancer therapy.

Enhancing Endothelial Cell Retention on ePTFE Constructs by siRNA-Mediated SHP-1 Gene Silencing

2011 ◽  
Vol 2 (1) ◽  
Author(s):  
Brandon J. Tefft ◽  
Adrian M. Kopacz ◽  
Wing Kam Liu ◽  
Shu Q. Liu
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Gene Silencing ◽  
Vascular Grafts ◽  
Sh2 Domain ◽  
Great Promise ◽  
Shear Stresses ◽  
Cell Retention ◽  
Stress Dependent

Polymeric vascular grafts hold great promise for vascular reconstruction, but the lack of endothelial cells renders these grafts susceptible to intimal hyperplasia and restenosis, precluding widespread clinical applications. The purpose of this study is to establish a stable endothelium on expanded polytetrafluoroethylene (ePTFE) membrane by small interfering RNA (siRNA)-induced suppression of the cell adhesion inhibitor SH2 domain-containing protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were treated with scrambled siRNA as a control or SHP-1 specific siRNA. Treated cells were seeded onto fibronectin-coated ePTFE scaffolds and exposed to a physiological range of pulsatile fluid shear stresses for 1 h in a variable-width parallel plate flow chamber. Retention of cells was measured and compared between various shear stress levels and between groups treated with scrambled siRNA and SHP-1 specific siRNA. HUVECs seeded on ePTFE membrane exhibited shear stress-dependent retention. Exposure to physiological shear stress (10 dyn/cm2) induced a reduction in the retention of scrambled siRNA treated cells from 100% to 85% at 1 h. Increased shear stress (20 dyn/cm2) further reduced retention of scrambled siRNA treated cells to 55% at 1 h. SHP-1 knockdown mediated by siRNA enhanced endothelial cell retention from approximately 60% to 85% after 1 h of exposure to average shear stresses in the range of 15–30 dyn/cm2. This study demonstrates that siRNA-mediated gene silencing may be an effective strategy for improving the retention of endothelial cells within vascular grafts.

scholarly journals Pharmacodynamic and Pharmacokinetic Properties of Full Phosphorothioate Small Interfering RNAs for Gene Silencing In Vivo

2020 ◽  
Author(s):  
Christian Berk ◽  
Gianluca Civenni ◽  
Yuluan Wang ◽  
Christian Steuer ◽  
Carlo V. Catapano ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Small Interfering Rnas ◽  
Pharmacokinetic Properties

An Automated High-Throughput Fluorescence In Situ Hybridization (FISH) Assay Platform for Use in the Identification and Optimization of siRNA-Based Therapeutics

2020 ◽  
pp. 247255522096004
Author(s):  
Hui H. Dou ◽  
Rommel Mallari ◽  
Andrew Pipathsouk ◽  
Amrita Das ◽  
Mei-Chu Lo
Keyword(s):  
In Situ Hybridization ◽  
Gene Silencing ◽  
Fluorescence In Situ Hybridization ◽  
High Throughput ◽  
Messenger Rna ◽  
Great Promise ◽  
Small Interfering Rnas ◽  
Response Curves ◽  
Vitro Assay

Since the revolutionary discovery of RNA interference (RNAi) more than 20 years ago, synthetic small interfering RNAs (siRNAs) have held great promise as therapeutic agents for treating human diseases by the specific knockdown of disease-causing gene products. To facilitate the development of siRNA therapeutics, a robust, high-throughput in vitro assay for measuring gene silencing is imperative during the initial siRNA lead sequence identification and, later, during the lead optimization with chemically modified siRNAs. There are several potential assays for measuring gene expression. Quantitative reverse transcription PCR (qRT-PCR) has been widely used to quantitate messenger RNA (mRNA). This method has a few disadvantages, however, such as the requirement for RNA isolation, complementary DNA (cDNA) generation, and PCR reaction, which are labor-intensive, limit the assay throughput, and introduce variability. We chose a high-content imaging assay, bDNA FISH, that combines the branched DNA (bDNA) technology with fluorescence in situ hybridization (FISH) to measure gene silencing by siRNAs because it is sensitive and robust with a short reagent procurement and assay development time. We also built a fully automated liquid-handling platform for executing bDNA FISH assays to increase throughput, and the system has a capacity of generating 192 concentration–response curves in a single run. We have successfully developed and executed the bDNA FISH assays for multiple targets using this automated platform to identify and optimize siRNA candidate molecules. Examples of the bDNA FISH assay for selected targets are presented.

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