RNA Interference by Short Hairpin RNAs Expressed in Vertebrate Cells

2004 ◽  
pp. 255-266 ◽  
Author(s):  
Gregory J. Hannon ◽  
Douglas S. Conklin
Keyword(s):  
Rna Interference ◽  
Short Hairpin ◽  
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.

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 Large-Scale, High-Throughput Validation of Short Hairpin RNA Sequences for RNA Interference

2006 ◽  
Vol 11 (3) ◽  
pp. 236-246 ◽  
Author(s):  
Laurence H. Lamarcq ◽  
Bradley J. Scherer ◽  
Michael L. Phelan ◽  
Nikolai N. Kalnine ◽  
Yen H. Nguyen ◽  
...  
Keyword(s):  
High Throughput ◽  
Large Scale ◽  
Strong Support ◽  
Gc Content ◽  
Rapid Identification ◽  
Hairpin Rna ◽  
Rna Sequences ◽  
Short Hairpin ◽  
Short Hairpin Rnas ◽  
Interfering Rna

A method for high-throughput cloning and analysis of short hairpin RNAs (shRNAs) is described. Using this approach, 464 shRNAs against 116 different genes were screened for knockdown efficacy, enabling rapid identification of effective shRNAs against 74 genes. Statistical analysis of the effects of various criteria on the activity of the shRNAs confirmed that some of the rules thought to govern small interfering RNA (siRNA) activity also apply to shRNAs. These include moderate GC content, absence of internal hairpins, and asymmetric thermal stability. However, the authors did not find strong support for positionspecific rules. In addition, analysis of the data suggests that not all genes are equally susceptible to RNAinterference (RNAi).

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