RNA interference: From gene silencing to gene-specific therapeutics

https://doi.org/10.1016/j.pharmthera.2005.03.004Get rights and content

Abstract

In the past 4 years, RNA interference (RNAi) has become widely used as an experimental tool to analyse the function of mammalian genes, both in vitro and in vivo. By harnessing an evolutionary conserved endogenous biological pathway, first identified in plants and lower organisms, double-stranded RNA (dsRNA) reagents are used to bind to and promote the degradation of target RNAs, resulting in knockdown of the expression of specific genes. RNAi can be induced in mammalian cells by the introduction of synthetic double-stranded small interfering RNAs (siRNAs) 21–23 base pairs (bp) in length or by plasmid and viral vector systems that express double-stranded short hairpin RNAs (shRNAs) that are subsequently processed to siRNAs by the cellular machinery. RNAi has been widely used in mammalian cells to define the functional roles of individual genes, particularly in disease. In addition, siRNA and shRNA libraries have been developed to allow the systematic analysis of genes required for disease processes such as cancer using high throughput RNAi screens. RNAi has been used for the knockdown of gene expression in experimental animals, with the development of shRNA systems that allow tissue-specific and inducible knockdown of genes promising to provide a quicker and cheaper way to generate transgenic animals than conventional approaches. Finally, because of the ability of RNAi to silence disease-associated genes in tissue culture and animal models, the development of RNAi-based reagents for clinical applications is gathering pace, as technological enhancements that improve siRNA stability and delivery in vivo, while minimising off-target and nonspecific effects, are developed.

Keywords

RNAi
siRNA
shRNA
Disease
Validation
Therapy

Abbreviations

AAV
adeno-associated virus
ABCA1
ATP-binding cassette, subfamily A, member 1
Bcl-2
B-cell chronic lymphocytic leukemia/lymphoma 2
BCR-ABL
breakpoint cluster region-protooncogene tyrosine-protein kinase ABL1 fusion protein
BMP4
bone morphogenetic protein 4
bp
base pairs
B-RAF
murine sarcoma viral (v-raf) oncogene homolog B1 oncogene
CCR5
chemokine (C–C motif) receptor 5
CD8
T-lymphocyte differentiation antigen T8/Leu-2
CML
chronic myelogenous leukemia
COPD
chronic obstructive pulmonary disease
Cre
causes recombination
CXCR4
chemokine (C–X–C motif), receptor 4
dsRNA
double-stranded RNA
EC
endothelial cell
EGFR
epidermal growth factor receptor
EpCAM
tumour-associated calcium signal transducer 1
ERK
extracellular signal regulated protein kinase
Fas
tumour necrosis factor receptor superfamily, member 6
HBV
hepatitis B virus
HCV
hepatitis C virus
HIV
human immunodeficiency virus
HMG-CoA
beta-hydroxy-beta-methylglutaryl coenzyme A
IFN
interferon
KC
keratinocyte-derived chemokine
K-Ras
c-Kirsten-ras protein
LacZ
β-galactosidase α-peptide
LNA
locked nucleic acid
LoxP
locus χ of crossover P1
LPS
lipopolysaccharide
MAPK
mitogen-activated protein kinase
MDR
multidrug-resistance protein
MIP2
macrophage inflammatory protein-2
miRNA
microRNAs
NF-κB
Nuclear factor-kappaB
PEG
polyethylene glycol
PEI
polyethyleneimine
PKR
dsRNA-dependent protein kinase
RasGAP
Ras GTPase-activating protein
RdDM
RNA-directed DNA methylation
RISC
RNA-induced silencing complex
RNAi
RNA interference
Rnase III
ribonuclease III
RSV
respiratory syncytial virus
SARS
severe acute respiratory syndrome
SCID
severe combined immunodeficiency
shRNA
short hairpin RNA
siRNA
small interfering RNA
S1P
sphingosine 1-phosphate
smad 1
mothers against decapentaplegic homolog 1
smad 7
mothers against decapentaplegic homolog 7
TGFα
transforming growth factor α
TGFβ
transforming growth factor β
TLR3
toll-like receptor 3
TNFα
tumour necrosis factor α
VEGF
vascular endothelial growth factor

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