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本帖最后由 细胞海洋 于 2011-6-25 11:08 编辑 * j# E' c( f: h* N$ t5 A
" j, Q' t. G9 B9 l# UPreface
+ f2 g) Z" A! k" c/ SLittle over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that
8 x! t8 E- ~& Q* }. ndouble-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode 2 B( ^ }$ W- Z
Caenorhabditis elegans (RNA interference, RNAi). This work converged with research in ( M: c9 C0 }/ Q: x/ a7 a
plants, in which related RNA-based silencing processes were known to exist. Ever since,
& f- I% Z. H3 q' _research in the feld has progressed at an astonishing rate, resulting in our appreciation of
9 \7 e; t8 y/ Z- d4 O( Fsmall silencing RNAs as central regulators of gene expression, as guards of genome integ-8 n+ i8 V' f0 h/ M$ i
rity, and as essential mediators of antiviral defense. The discovery that synthetic small * Y S$ `4 `: s3 I- R; S% f" n' P
interfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-+ n" W1 \+ _) E5 D/ R. C/ S
leagues in 2001, has further boosted the development of novel therapeutics and experi-
; }9 M/ V7 _: m2 k; f, G5 G# [mental tools based on RNAi technology.
, x+ X& I7 j' t' j6 O; B$ v# f0 ~5 UViruses and RNAi share an intricate relationship at many levels. Early work in plants
, r; c+ w5 d3 x# q% P: a7 w0 ]indicated that viruses can be both inducers and targets of RNA-based post-transcriptional
4 D) w, i+ x' M% R6 Wgene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as : Q1 m3 i' \1 f; q
an antiviral defense mechanism is now well-established in plants and other organisms,
9 i$ [$ ]" n2 G. v! kincluding insects. In vertebrates, viruses also interact with a related RNA silencing mecha-2 a/ I& X) F4 n; L" T/ @
nism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set
8 u% V9 X# {4 R7 w; {- r# C, x9 wof miRNAs, by which they regulate viral or host gene expression and modify, for example,
% D, m: w; [6 z& cthe transition from latent to lytic infection and the recognition of infected cells by the host , b' C+ d6 h& A6 K: q6 X2 B% M; m
immune system. Furthermore, cellular miRNAs likely regulate expression of many genes
z: Z' c$ d* R6 N$ L) v8 Kthat are important for virus biology, but they have been suggested to directly target viral ' Y: K3 F% M( L
RNA as well.) J+ T% o& L$ k5 }" B/ f
The therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is
5 v$ x' n; U/ T8 wnow clear that replication of many, if not all, mammalian viruses can be suppressed by 4 P/ i% V: q. D7 t& I' j
RNAi in cell culture. While these results have raised considerable optimism about the ' @ @. n3 C& ^+ n
potential of RNAi-based drugs, important hurdles remain, including issues related to the % Q( m; P8 j3 O3 v6 e5 ]! z R
delivery and stability of siRNAs and the risk of viral escape.
' z ]) m; s$ ~+ I+ `From this brief overview it will be apparent that a great — and increasing — number
! b% y; Q( G8 Z8 J; K9 C5 fof tools and techniques are available for those interested in the interface of viruses and * |, _* j) p3 d6 c
RNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-0 v2 c1 N( k! A# a7 G) X
cols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for
! h; t Y( X+ j0 j. P8 S1 h3 c3 C0 L9 ]the development and optimization of RNAi-based antiviral drugs. As RNAi is a central
( D' K r; N# d: Q! fregulatory mechanism in the cell, the methods in this volume can also be applied out of
- g0 \/ d2 | L% k4 p( pthe context of a virus infection. In the established tradition of the Methods in Molecular 4 R4 }# [9 H, b. V, d2 p
Biology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-# r' B& Z* ~5 H$ u/ U
by-step protocols and extra tools and tricks that should be useful to those new to the feld
, E! x. A; i5 F0 Hand experienced scientists alike.7 f4 a' }% y% d8 U
This volume consists of fve parts. Part 1 reviews important basic concepts in the feld
- _+ H* N7 c9 \2 |' c3 bof antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of ^0 H0 Y' d C! W' R2 r
small silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-" e9 {5 W: H9 G, o T' L$ `4 D
ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
2 e% v7 l" H0 J( m% A" q% Qexpression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide ( C8 e* o+ X2 C6 C/ c/ B
RNAi approaches for the identifcation of factors involved in virus replication, which may
3 ]0 [( P9 O- r8 h/ h8 t' mrepresent novel targets for antiviral therapy.% t& H# V% F4 o# i0 x: l& a% a' q) F
I am grateful to all authors for providing their outstanding contributions and to John
8 T5 M: M% k. B" v% M! t: OWalker for guidance while editing this volume. I thank members of my lab, especially
' x& ~/ U2 ?) }: sWalter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus- P6 |) J! Q6 J+ r
sions. I am thankful to Raul Andino for having been a great mentor and for introducing
4 }1 E, s8 v0 v& d7 ~# @* k1 [me to this exciting feld of research. Finally, I would like to apologize for doing little jus-
- @5 k& {3 r3 ]* H! O0 u" ztice to the seminal work in plants; space limitations forced me to focus this volume on the
1 E$ { L. w- d$ Y( A4 e9 canimal system.
1 t: M4 F( G+ H; J! {4 _Nijmegen, The Netherlands Ronald P. van Rij |
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发表于 2011-6-25 10:44
