scholarly journals Enhancing Chemotherapy by RNA Interference

2020 ◽  
Vol 1 (2) ◽  
pp. 64-81
Author(s):  
Shuwen Cao ◽  
Chunhao Lin ◽  
Shunung Liang ◽  
Chee Hwee Tan ◽  
Phei Er Saw ◽  
...  
Keyword(s):  
Anticancer Drugs ◽  
Sequential Therapy ◽  
Sirna Treatment ◽  
Rnai Technology ◽  
New Strategy ◽  
Multiple Challenges ◽  
Sirna Therapy ◽  
Interfering Rna ◽  
Target Side ◽  
Specific Mrna

Abstract Small interfering RNA (siRNA) has shown tremendous potential for treating human diseases in the past decades. siRNA can selectively silence a pathological pathway through the targeting and degradation of a specific mRNA, significantly reducing the off-target side effects of anticancer drugs. However, the poor pharmacokinetics of RNA significantly restricted the clinical use of RNAi technology. In this review, we examine in-depth the siRNA therapeutics currently in preclinical and clinical trials, multiple challenges faced in siRNA therapy, feasibility of siRNA treatment with anticancer drugs in combined with siRNA in nanoparticles or modified to be parental drugs, sequential therapy of siRNA treatment prior to drug treatment with siRNA and drugs loaded in nanoparticles. We focused on the combinatorial activation of apoptosis by different pathways, namely Bcl-2, survivin, and Pgp protein. Taken together, this review would serve to establish the pathway of effective and efficient combination therapy of siRNA and drugs as a new strategy.

scholarly journals RNAi Technology in Fish and Shellfish- Status and Prospects: A Review

2020 ◽  
Author(s):  
Ubaid Qayoom ◽  
Zahoor Mushtaq
Keyword(s):  
Small Interfering Rna ◽  
Therapeutic Strategy ◽  
Fish Farming ◽  
Economic Losses ◽  
Rnai Technology ◽  
The Status ◽  
Interfering Rna ◽  
Fish And Shellfish ◽  
Specific Mrna

Ribonucleic acid interference (RNAi), a valuable tool for manipulating gene functionality in the laboratory, has also emerged as a powerful tool to suppress infection or replication of many pathogens that cause severe economic losses in fish farming. By taking advantage of the cell’s endogenous RNAi apparatus, small interfering RNA of ~21-22 bp can be introduced into cells to induce target specific mRNA degradation. With the growing appreciation for the potential of RNAi technology, the diversity in vivo relevance to aquaculture is seemingly vast. Studies in the future should address the hurdles like delivery strategy stability and degradation of RNAi therapeutic molecule by nucleases in aquatic animals. In this article, we review the literature in the field of RNAi technology in aquaculture, summarize the status and prospects, which may open doors to its applicability potential as a therapeutic strategy to modulate host-pathogen interactions and inspire further trials.

Single and Combinational siRNA Therapy of Cancer Cells: Probing Changes in Targeted and Nontargeted Mediators after siRNA Treatment

2016 ◽  
Vol 13 (12) ◽  
pp. 4116-4128 ◽  
Author(s):  
Hamidreza Montazeri Aliabadi ◽  
Parvin Mahdipoor ◽  
Marco Bisoffi ◽  
Judith C. Hugh ◽  
Hasan Uludağ
Keyword(s):  
Cancer Cells ◽  
Sirna Treatment ◽  
Sirna Therapy

scholarly journals BiP Internal Ribosomal Entry Site Activity Is Controlled by Heat-Induced Interaction of NSAP1

2006 ◽  
Vol 27 (1) ◽  
pp. 368-383 ◽  
Author(s):  
Sungchan Cho ◽  
Sung Mi Park ◽  
Tae Don Kim ◽  
Jong Heon Kim ◽  
Kyong-Tai Kim ◽  
...  
Keyword(s):  
Heat Stress ◽  
Protein A ◽  
Internal Ribosomal Entry Site ◽  
Stress Conditions ◽  
Entry Site ◽  
Sirna Treatment ◽  
Nontranslated Region ◽  
Ires Element ◽  
Ribosomal Entry ◽  
Interfering Rna

ABSTRACT TheBiP protein, a stress response protein, plays an important role in the proper folding and assembly of nascent protein and in the scavenging of misfolded proteins in the endoplasmic reticulum lumen. Translation of BiP is directed by an internal ribosomal entry site (IRES) in the 5′ nontranslated region of the BiP mRNA. BiP IRES activity increases when cells are heat stressed. Here we report that NSAP1 specifically enhances the IRES activity of BiP mRNA by interacting with the IRES element. Overexpression of NSAP1 in 293T cells increased the IRES activity of BiP mRNA, whereas knockdown of NSAP1 by small interfering RNA (siRNA) reduced the IRES activity of BiP mRNA. The amount of NSAP1 bound to the BiP IRES increased under heat stress conditions, and the IRES activity of BiP mRNA was increased. Moreover, the increase in BiP IRES activity with heat treatment was not observed in cells lacking NSAP1 after siRNA treatment. BiP mRNAs were redistributed from the heavy polysome to the light polysome in NSAP1 knockdown cells. Together, these data indicate that NSAP1 modulates IRES-dependent translation of BiP mRNA through an RNA-protein interaction under heat stress conditions.

High-Content Screening with siRNA Optimizes a Cell Biological Approach to Drug Discovery: Defining the Role of P53 Activation in the Cellular Response to Anticancer Drugs

2004 ◽  
Vol 9 (7) ◽  
pp. 557-568 ◽  
Author(s):  
Kenneth A. Giuliano ◽  
Yih-Tai Chen ◽  
D. Lansing Taylor
Keyword(s):  
Cell Cycle ◽  
Drug Discovery ◽  
Anticancer Drugs ◽  
Cellular Response ◽  
P53 Protein ◽  
High Content Screening ◽  
M Cell ◽  
Model Application ◽  
Rnai Technology ◽  
Blocking Activity

Deciphering the effects of compounds on molecular events within living cells is becoming an increasingly important component of drug discovery. In a model application of the industrial drug discovery process, the authors profiled a panel of 22 compounds using hierarchical cluster analysis of multiparameter high-content screening measurements from nearly 500,000 cells per microplate. RNAi protein knockdown methodology was used with high-content screening to dissect the effects of 2 anticancer drugs on multiple target activities. Camptothecin activated p53 in A549 lung carcinoma cells pretreated with scrambled siRNA, exhibited concentration-dependent cell cycle blocks, and induced moderate microtubule stabilization. Knockdown of camptothecin-induced p53 protein expression with p53 siRNA inhibited the G1/S blocking activity of the drug and diminished its microtubule-stabilizing activity. Paclitaxel activated p53 protein at low concentrations but exhibited G2/M cell cycle blocking activity at higher concentrations where microtubules were stabilized. In cells treated with p53 siRNA, paclitaxel failed to activate p53 protein, but the knockdown did not have a significant effect on the ability of paclitaxel to stabilize microtubules or induce a G2/M cell cycle block. Thus, this model application of the use of RNAi technology within the context of high-content screening shows the potential to provide massive amounts of combinatorial cell biological information on the temporal and spatial responses that cells mount to treatment by promising therapeutic candidates.

Abstract 4386: CETSA as a new strategy to understand efficacy, adverse effects and resistance development of anticancer drugs

2016 ◽  
Author(s):  
Pär Nordlund ◽  
Sara Lööf ◽  
Henritte Laursen ◽  
Anette Öberg ◽  
Johan Lengqvist ◽  
...  
Keyword(s):  
Adverse Effects ◽  
Anticancer Drugs ◽  
Resistance Development ◽  
New Strategy

scholarly journals Electroporation Knows No Boundaries

2015 ◽  
Vol 20 (8) ◽  
pp. 932-942 ◽  
Author(s):  
Christin Luft ◽  
Robin Ketteler
Keyword(s):  
Cell Lines ◽  
High Throughput ◽  
High Throughput Screening ◽  
High Efficiency ◽  
Target Identification ◽  
Cell Types ◽  
Rnai Technology ◽  
Drug Target Identification ◽  
Interfering Rna ◽  
Calcium Phosphate Precipitation

The discovery of RNA interference (RNAi) has enabled several breakthrough discoveries in the area of functional genomics. The RNAi technology has emerged as one of the major tools for drug target identification and has been steadily improved to allow gene manipulation in cell lines, tissues, and whole organisms. One of the major hurdles for the use of RNAi in high-throughput screening has been delivery to cells and tissues. Some cell types are refractory to high-efficiency transfection with standard methods such as lipofection or calcium phosphate precipitation and require different means. Electroporation is a powerful and versatile method for delivery of RNA, DNA, peptides, and small molecules into cell lines and primary cells, as well as whole tissues and organisms. Of particular interest is the use of electroporation for delivery of small interfering RNA oligonucleotides and clustered regularly interspaced short palindromic repeats/Cas9 plasmid vectors in high-throughput screening and for therapeutic applications. Here, we will review the use of electroporation in high-throughput screening in cell lines and tissues.

scholarly journals The Melanocortin 2 Receptor Accessory Protein Exists as a Homodimer and Is Essential for the Function of the Melanocortin 2 Receptor in the Mouse Y1 Cell Line

Endocrinology ◽  
2007 ◽  
Vol 149 (4) ◽  
pp. 1935-1941 ◽  
Author(s):  
Sadani N. Cooray ◽  
Isabel Almiro Do Vale ◽  
Kit-Yi Leung ◽  
Tom R. Webb ◽  
J. Paul Chapple ◽  
...  
Keyword(s):  
Sodium Dodecyl ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Accessory Protein ◽  
Rnai Technology ◽  
Accessory Factor ◽  
Acth Receptor ◽  
Sds Page ◽  
Lower Mobility ◽  
Interfering Rna

The ACTH receptor [melanocortin 2 receptor (MC2R)] gene produces a functional receptor only when transfected into cells of adrenocortical origin, implying that it may require an adrenal-specific accessory factor. Recently we showed that the MC2R accessory protein (MRAP) is essential for the cell surface expression of the MC2R in such models. Using RNA interference (RNAi) technology, we have further explored the action of MRAP in the functioning of the MC2R in Y1 mouse adrenocortical cells that endogenously express MRAP and MC2R. We created stable cell lines expressing mouse MRAP short hairpin RNA (shRNAs) by transfecting cells with an expression vector containing the MRAP small interfering RNA sequence. The knockdown of MRAP resulted in a reduction in MC2R signaling. The overexpression of a mouse MRAP-Flag construct did not restore the expression of MRAP due to its degradation by the mouse shRNAs. The introduction of human MRAP that is resistant to silencing by mouse MRAP shRNAs resulted in the rescue of the MC2R signaling. MRAP migrates on SDS-PAGE with markedly lower mobility than predicted for a 14.1-kDa protein. Coimmunoprecipitation and mass spectroscopy suggests that MRAP exists as a homodimer that is resistant to dissociation by sodium dodecyl sulfate and reducing agents.

scholarly journals Small Interfering RNA- Modern Approach for Intervening Pathological Conditions

2021 ◽  
Vol 2 (2) ◽  
pp. 16-19
Author(s):  
Shazia Choudhary ◽  
Sheeba Murad ◽  
Sana Gul ◽  
Hayat Khan ◽  
Samra Khalid ◽  
...  
Keyword(s):  
Rna Interference ◽  
Small Interfering Rna ◽  
Sirna Delivery ◽  
Rnai Pathway ◽  
Modern Approach ◽  
Rna Molecules ◽  
Rnai Technology ◽  
Efficient Delivery ◽  
Biological Discovery ◽  
Interfering Rna

RNA interference (RNAi) refers to the inhibition of gene expression by small double-stranded RNA molecules. This technology can prove to be a breakthrough biological discovery of the decade as it has the potential to revolutionize the field of therapeutics. RNA interference (RNAi) through small interfering RNA (siRNA) is currently being evaluated for its efficacy to be used in therapeutics as well as prophylactic strategies. Many studies are being conducted across the globe to optimize the siRNA delivery systems (in terms of safe, stable and efficient delivery) in various disorders. There are a number of diseases such as autoimmune diseases, cancer associated pathological changes, bacterial and viral induced disorders, where RNAi pathway can be explored and RNAi technology can be used as a tool to intervene such abnormalities. This review is an effort to review latest advancements in the field of siRNA based therapy development and the pits and falls generally encountered in the use of this technology.

Successful silencing of plasminogen activator inhibitor-1 in human vascular endothelial cells using small interfering RNA

2006 ◽  
Vol 95 (05) ◽  
pp. 857-864 ◽  
Author(s):  
Anneke Hecke ◽  
Hilary Brooks ◽  
Matthieu Meryet-Figuière ◽  
Stephanie Minne ◽  
Stavros Konstantinides ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Small Interfering Rna ◽  
Vascular Endothelial Cells ◽  
Plasminogen Activator Inhibitor ◽  
Vascular Endothelial ◽  
Sirna Treatment ◽  
Interfering Rna ◽  
Activator Inhibitor ◽  
Pai 1

SummaryClinical as well as experimental evidence suggests that vascular overexpression of plasminogen activator inhibitor (PAI)-1, the primary physiological inhibitor of both urokinase and tissuetype plasminogen activator, may be involved in the pathophysiology of atherosclerosis and cardiovascular disease. We investigated the feasibility, efficacy and functional effects of PAI-1 gene silencing in human vascular endothelial cells using small interfering RNA. Double-stranded 21 bp-RNA molecules targeted at sequences within the human PAI-1 gene were constructed. Successful siRNA transfection of HUVEC was confirmed using fluorescence microscopy and flow cytometry. One of five candidate siRNA sequences reduced PAI-1 mRNA and protein in a concentrationand time-dependent manner. Suppression of PAI-1 mRNA was detected up to 72 hours after transfection. Moreover, siRNA treatment reduced the activity of PAI-1 released from HUVEC, and prevented the oxLDL- or LPS-induced upregulation of PAI-1 secretion. Importantly, siRNA treatment did not affect the expression of other endothelial-cell markers. Moreover, downregulation of PAI-1 significantly enhanced the ability of endothelial cells to adhere to vitronectin, and this effect could be reversed upon addition of recombinant PAI-1. SiRNAmediated reduction of PAI-1 expression may be a promising strategy for dissecting the effects of PAI-1 on vascular homeostasis.

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