RNA Interference Therapy for Machado–Joseph Disease: Long-Term Safety Profile of Lentiviral Vectors Encoding Short Hairpin RNAs Targeting Mutant Ataxin-3

2019 ◽  
Vol 30 (7) ◽  
pp. 841-854 ◽  
Author(s):  
Clévio Nóbrega ◽  
José Miguel Codêsso ◽  
Liliana Mendonça ◽  
Luís Pereira de Almeida
Keyword(s):  
Rna Interference ◽  
Safety Profile ◽  
Lentiviral Vectors ◽  
Short Hairpin ◽  
Machado Joseph Disease ◽  
Short Hairpin Rnas

scholarly journals A versatile approach to multiple gene RNA interference using microRNA-based short hairpin RNAs

2007 ◽  
Vol 8 (1) ◽  
pp. 98 ◽  
Author(s):  
Xiaocui Zhu ◽  
Leah A Santat ◽  
Mi Chang ◽  
Jamie Liu ◽  
Joelle R Zavzavadjian ◽  
...  
Keyword(s):  
Rna Interference ◽  
Multiple Gene ◽  
Short Hairpin ◽  
Short Hairpin Rnas

scholarly journals Synthetic Pre-miRNA-Based shRNA as Potent RNAi Triggers

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Kazuya Terasawa ◽  
Kazuharu Shimizu ◽  
Gozoh Tsujimoto
Keyword(s):  
Biological Activity ◽  
Rna Interference ◽  
Gene Function ◽  
Small Interfering Rnas ◽  
Therapeutic Applications ◽  
Short Hairpin ◽  
Short Hairpin Rnas ◽  
Interferon Responses ◽  
Rnai Activity ◽  
Target Effects

RNA interference (RNAi) is a powerful tool for studying gene function owing to the ease with which it can selectively silence genes of interest, and it has also attracted attention because of its potential for therapeutic applications. Chemically synthesized small interfering RNAs (siRNAs) and DNA vector-based short hairpin RNAs (shRNAs) are now widely used as RNAi triggers. In contrast to expressed shRNAs, the use of synthetic shRNAs is limited. Here we designed shRNAs modeled on a precursor microRNA (pre-miRNA) and evaluated their biological activity. We demonstrated that chemically synthetic pre-miRNA-based shRNAs have more potent RNAi activity than their corresponding siRNAs and found that their antisense strands are more efficiently incorporated into the RNA-induced silencing complex. Although greater off-target effects and interferon responses were induced by shRNAs than by their corresponding siRNAs, these effects could be overcome by simply using a lower concentration or by optimizing and chemically modifying shRNAs similar to synthetic siRNAs. These are challenges for the future.

scholarly journals Transfer and Expression of Small Interfering RNAs in Mammalian Cells Using Lentiviral Vectors

Acta Naturae ◽  
2013 ◽  
Vol 5 (2) ◽  
pp. 7-18 ◽  
Author(s):  
T. D. Lebedev ◽  
P. V. Spirin ◽  
V. S. Prassolov
Keyword(s):  
Rna Interference ◽  
Mammalian Cells ◽  
Lentiviral Vectors ◽  
Cell Delivery ◽  
Small Interfering Rnas ◽  
Shrna Expression ◽  
Widespread Application ◽  
Efficient Gene ◽  
Interfering Rna

RNA interference is a convenient tool for modulating gene expression. The widespread application of RNA interference is made difficult because of the imperfections of the methods used for efficient target cell delivery of whatever genes are under study. One of the most convenient and efficient gene transfer and expression systems is based on the use of lentiviral vectors, which direct the synthesis of small hairpin RNAs (shRNAs), the precursors of siRNAs. The application of these systems enables one to achieve sustainable and long-term shRNA expression in cells. This review considers the adaptation of the processing of artificial shRNA to the mechanisms used by cellular microRNAs and simultaneous expression of several shRNAs as potential approaches for producing lentiviral vectors that direct shRNA synthesis. Approaches to using RNA interference for the treatment of cancer, as well as hereditary and viral diseases, are under active development today. The improvement made to the methods for constructing lentiviral vectors and the investigation into the mechanisms of processing of small interfering RNA allow one to now consider lentiviral vectors that direct shRNA synthesis as one of the most promising tools for delivering small interfering RNAs.

Artificial miRNAs are potential gene therapy tools, especially for incurable monogenic disorders

2021 ◽  
Author(s):  
Moataz Dowaidar
Keyword(s):  
Gene Therapy ◽  
Brain Surgery ◽  
Native Protein ◽  
Intravenous Injections ◽  
Monogenic Disorders ◽  
Monogenic Diseases ◽  
Short Hairpin ◽  
The Cost ◽  
Short Hairpin Rnas

Well-designed artificial miRNAs (amiRNAs) are as effective as short hairpin RNAs (shRNAs) but produce 10–80 times less siRNA. They enable long-term silencing and are safer than other RNAi triggers. They are suitable instruments for gene therapy techniques, especially for incurable monogenic diseases. In clinical studies, stereotactic injection of AAV5 directly into the striatum is the most effective approach. Intravenous injections would not only make patients more comfortable, but would also reduce the cost of complex brain surgery. In terms of structure, biogenesis, and expression levels, Ami RNAs are more "natural" than other gene therapy methods. They also utilise the cell's native protein machinery and do not produce irreversible alterations, unlike genome editing technologies. The amount of time spent on a technology determines its level of progression. ASOs have an edge in this regard, as seen by the number of authorized medicines. Perhaps RNAi is just around the corner.

Alu-linked hairpins efficiently mediate RNA interference with less toxicity than do H1-expressed short hairpin RNAs

2006 ◽  
Vol 349 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Stephen L. Gasior ◽  
Melanie Palmisano ◽  
Prescott L. Deininger
Keyword(s):  
Rna Interference ◽  
Short Hairpin ◽  
Short Hairpin Rnas

scholarly journals Gene knockdown via electroporation of short hairpin RNAs in embryos of the marine hydroid Hydractinia symbiolongicarpus

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Gonzalo Quiroga-Artigas ◽  
Alexandrea Duscher ◽  
Katelyn Lundquist ◽  
Justin Waletich ◽  
Christine E. Schnitzler
Keyword(s):  
Gene Function ◽  
Marine Invertebrates ◽  
Endogenous Gene ◽  
Successful Model ◽  
Hydractinia Symbiolongicarpus ◽  
Endogenous Genes ◽  
Short Hairpin ◽  
Visual Confirmation ◽  
Short Hairpin Rnas

Abstract Analyzing gene function in a broad range of research organisms is crucial for understanding the biological functions of genes and their evolution. Recent studies have shown that short hairpin RNAs (shRNAs) can induce gene-specific knockdowns in two cnidarian species. We have developed a detailed, straightforward, and scalable method to deliver shRNAs into fertilized eggs of the hydrozoan cnidarian Hydractinia symbiolongicarpus via electroporation, yielding effective gene-targeted knockdowns that can last throughout embryogenesis. Our electroporation protocol allows for the transfection of shRNAs into hundreds of fertilized H. symbiolongicarpus eggs simultaneously with minimal embryo death and no long-term harmful consequences on the developing animals. We show RT-qPCR and detailed phenotypic evidence of our method successfully inducing effective knockdowns of an exogenous gene (eGFP) and an endogenous gene (Nanos2), as well as knockdown confirmation by RT-qPCR of two other endogenous genes. We also provide visual confirmation of successful shRNA transfection inside embryos through electroporation. Our detailed protocol for electroporation of shRNAs in H. symbiolongicarpus embryos constitutes an important experimental resource for the hydrozoan community while also serving as a successful model for the development of similar methods for interrogating gene function in other marine invertebrates.

Sign in / Sign up

Export Citation Format

Share Document