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本帖最后由 细胞海洋 于 2011-6-25 11:08 编辑
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' o+ w: d. n6 R% VPreface' g6 m! }* Y3 P j' i1 O
Little over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that " `. D) M, L% j
double-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode ) S; N% j! O" K
Caenorhabditis elegans (RNA interference, RNAi). This work converged with research in $ t. E1 N2 u; I& L ]0 R, f
plants, in which related RNA-based silencing processes were known to exist. Ever since, 9 D7 w: T" f! q& s! {
research in the feld has progressed at an astonishing rate, resulting in our appreciation of
% Q! z: j7 _7 K0 h" xsmall silencing RNAs as central regulators of gene expression, as guards of genome integ-
4 r! M& Y9 H0 f/ m$ Xrity, and as essential mediators of antiviral defense. The discovery that synthetic small \! v& H n, P9 Q8 Y* p
interfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-
8 ] l# @" o( o) ^" j0 [ T* ^2 d& Tleagues in 2001, has further boosted the development of novel therapeutics and experi-
- v3 z' p! m8 amental tools based on RNAi technology.. g8 [3 v6 o% x7 n5 j
Viruses and RNAi share an intricate relationship at many levels. Early work in plants
/ ~9 p# h5 O1 Z; I' R7 Vindicated that viruses can be both inducers and targets of RNA-based post-transcriptional , n( y: q7 X1 l6 {# C* m
gene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as
& A) ^6 B: s9 v, m7 X$ ]" \an antiviral defense mechanism is now well-established in plants and other organisms, : j" F- S+ o% c/ x9 k3 b
including insects. In vertebrates, viruses also interact with a related RNA silencing mecha-4 ]7 k) `8 E- U. J
nism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set ' ^1 U; @# @5 W( @; P u+ A3 B
of miRNAs, by which they regulate viral or host gene expression and modify, for example,
; i" b2 _: Y" P4 [7 x& kthe transition from latent to lytic infection and the recognition of infected cells by the host ( [3 U) x G" Q0 m
immune system. Furthermore, cellular miRNAs likely regulate expression of many genes 8 V) s) S& ~ \
that are important for virus biology, but they have been suggested to directly target viral
5 C& s' s, ~# U2 o. g7 w# n8 Q- Z8 e( NRNA as well.7 Y# X- {; {8 v3 I0 [' ~( M
The therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is
, R' E) m, w( M' {, A( Vnow clear that replication of many, if not all, mammalian viruses can be suppressed by " a- d# m1 t5 c
RNAi in cell culture. While these results have raised considerable optimism about the ) b: [# l) \7 u
potential of RNAi-based drugs, important hurdles remain, including issues related to the + M( B1 e4 x: X7 ]! g
delivery and stability of siRNAs and the risk of viral escape.! U, Y! S2 X1 _0 v1 O
From this brief overview it will be apparent that a great — and increasing — number
8 Q' \, f# C% b! b( s0 h9 W: m9 Lof tools and techniques are available for those interested in the interface of viruses and ( B( U4 Z) t; v; b& O8 L% M6 k$ q' X S
RNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-! a2 Z" }4 Z/ U7 A* |; p- P" B
cols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for / O/ H! Z# x$ }9 r S
the development and optimization of RNAi-based antiviral drugs. As RNAi is a central / Z0 D3 U) L0 W) b/ C$ D6 Y
regulatory mechanism in the cell, the methods in this volume can also be applied out of
: c4 \$ r1 B0 Nthe context of a virus infection. In the established tradition of the Methods in Molecular
9 P9 d, j, B6 _ S% z- |; @Biology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-1 A/ F3 U; ]( a* T) j7 h
by-step protocols and extra tools and tricks that should be useful to those new to the feld
: Q: E2 g2 ?1 Sand experienced scientists alike.; h q# @7 `9 L8 o- b
This volume consists of fve parts. Part 1 reviews important basic concepts in the feld
5 ]1 j7 {% O5 R* [3 tof antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of
( U& }/ L( D2 N1 v0 Tsmall silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-
2 o( I1 m1 l0 t9 ^ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
& N$ S: F0 E W7 Qexpression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide # J6 q% w' I: p) t! X$ d+ O$ A
RNAi approaches for the identifcation of factors involved in virus replication, which may
# H7 d1 C, j* V$ ?) i( [% erepresent novel targets for antiviral therapy.! H& y2 g! J" W6 r, x, ~) M& X; g: a
I am grateful to all authors for providing their outstanding contributions and to John
* C) q) Z4 q0 ^( J, A0 [, fWalker for guidance while editing this volume. I thank members of my lab, especially
$ {) Q; h$ s+ H/ RWalter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-2 ~- k M! i, h4 h m
sions. I am thankful to Raul Andino for having been a great mentor and for introducing
/ x* X ]8 v7 X5 Z5 Nme to this exciting feld of research. Finally, I would like to apologize for doing little jus-
9 E* V2 L4 a0 dtice to the seminal work in plants; space limitations forced me to focus this volume on the
4 L) O2 L0 p* ?# {animal system.8 _7 Q3 [$ I, e Q# @ b
Nijmegen, The Netherlands Ronald P. van Rij |
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发表于 2011-6-25 10:44
发表于 2011-6-25 11:09
