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本帖最后由 细胞海洋 于 2011-6-25 11:08 编辑 ( c Q2 }: T$ ] I* B; q
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Preface
! r. t# p% ?( W! e) eLittle over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that * m" S7 p( W! P8 T
double-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode
! m" \+ Q6 Y" B/ E9 ACaenorhabditis elegans (RNA interference, RNAi). This work converged with research in 8 K+ v, Q# x) J" D
plants, in which related RNA-based silencing processes were known to exist. Ever since, ; T( k6 K& Z9 C! q9 \, Q
research in the feld has progressed at an astonishing rate, resulting in our appreciation of
6 }1 i3 M# }& L" Xsmall silencing RNAs as central regulators of gene expression, as guards of genome integ-
' a v9 e* H/ R8 j, hrity, and as essential mediators of antiviral defense. The discovery that synthetic small
/ k' @2 g5 l! W# d% i4 O9 Y T; Ninterfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-$ U& p5 G, \# O
leagues in 2001, has further boosted the development of novel therapeutics and experi-
3 b3 i; G4 `5 L4 L9 \mental tools based on RNAi technology.
! I: t8 K% l. IViruses and RNAi share an intricate relationship at many levels. Early work in plants ; ~ ?+ X2 I; I5 d+ k3 ~9 t ?' m
indicated that viruses can be both inducers and targets of RNA-based post-transcriptional 9 o+ O& M* g. C, A; d
gene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as
! F% ?3 h, t0 T( `' Uan antiviral defense mechanism is now well-established in plants and other organisms,
; ^3 V M% J! c7 D: Iincluding insects. In vertebrates, viruses also interact with a related RNA silencing mecha-9 s7 T0 j( ?7 w$ c4 k
nism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set
' V1 F- B. Y5 \6 H5 n% e, Tof miRNAs, by which they regulate viral or host gene expression and modify, for example,
1 N3 m, i, e" ]! `( Ethe transition from latent to lytic infection and the recognition of infected cells by the host
" w0 N& h8 p/ N- }/ b4 ?immune system. Furthermore, cellular miRNAs likely regulate expression of many genes 4 T5 c/ D# ~& n4 d8 M/ ^' C j9 g
that are important for virus biology, but they have been suggested to directly target viral
5 g6 D+ `3 }* m! Y/ [# B( w2 G9 \RNA as well. u8 [+ u6 P% i; C( f9 d' y" r) W# K
The therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is 7 l0 A* K* w" W" s+ G. `
now clear that replication of many, if not all, mammalian viruses can be suppressed by & l5 t$ t: ?. x5 ]6 o+ S4 K
RNAi in cell culture. While these results have raised considerable optimism about the
+ t' g& Q! H! p" @6 F/ ]/ F/ v) Gpotential of RNAi-based drugs, important hurdles remain, including issues related to the ! r- w# c; _. c8 W w" M! M
delivery and stability of siRNAs and the risk of viral escape.
, E* P& g. t! Q! MFrom this brief overview it will be apparent that a great — and increasing — number
+ W) W3 v2 g( a: P3 _2 q+ f9 O9 q9 Aof tools and techniques are available for those interested in the interface of viruses and ( \9 l; J7 m D( P
RNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-. \" g/ g5 K6 `8 L4 I
cols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for
" d* V8 n; ^$ O; [1 r, bthe development and optimization of RNAi-based antiviral drugs. As RNAi is a central 9 L a! Y$ [9 i& p( J, m8 G3 s8 v
regulatory mechanism in the cell, the methods in this volume can also be applied out of / q, ? S& k; \: K; u1 v
the context of a virus infection. In the established tradition of the Methods in Molecular : j/ t* k6 m7 g5 ^
Biology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-
- g1 [9 H$ ]6 T Z9 F7 W1 dby-step protocols and extra tools and tricks that should be useful to those new to the feld
" p9 P1 y4 e( Rand experienced scientists alike.3 G" g" D# v) @5 V
This volume consists of fve parts. Part 1 reviews important basic concepts in the feld 8 {* P8 q6 [" Z/ b
of antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of / m8 e: O, e3 l7 d! @
small silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-3 e8 o( v$ v V
ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design, ( d3 i) {1 I* T* `/ D! ?, K8 m
expression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide
: A A5 o. w9 m( k3 g; \5 r0 R% cRNAi approaches for the identifcation of factors involved in virus replication, which may
+ q9 S' |( z9 m% g" F- ~represent novel targets for antiviral therapy.0 T7 u k# ]& `" ^/ J4 @5 s. j7 |
I am grateful to all authors for providing their outstanding contributions and to John
; _9 F2 D% g( r; oWalker for guidance while editing this volume. I thank members of my lab, especially + U/ `0 r4 H2 G* b
Walter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-& }/ j/ l6 c f+ m
sions. I am thankful to Raul Andino for having been a great mentor and for introducing ' q" x( \/ P; i6 z V( {
me to this exciting feld of research. Finally, I would like to apologize for doing little jus-8 B& d- S: O* X* `! O! o
tice to the seminal work in plants; space limitations forced me to focus this volume on the
1 w7 Z5 g. j& y" P$ n/ _' _animal system.
" ]4 @* z' c @8 l7 T( ^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
