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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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) t: m$ l9 }, B3 [2 h7 q# MPreface
@7 T4 a7 _' ^& y+ ALittle over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that
! `3 p& s& n# `* W7 a$ kdouble-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode 2 P8 v+ W2 a" {; o( m4 N, ?$ R
Caenorhabditis elegans (RNA interference, RNAi). This work converged with research in
2 m# w2 e& P4 n# P4 M; Fplants, in which related RNA-based silencing processes were known to exist. Ever since,
- G, k7 z0 Q3 {% y: j; Y9 Cresearch in the feld has progressed at an astonishing rate, resulting in our appreciation of ; Y5 J% F$ I C" x( }- `+ s
small silencing RNAs as central regulators of gene expression, as guards of genome integ-
6 D3 i6 e( W/ o1 `/ ~4 U/ erity, and as essential mediators of antiviral defense. The discovery that synthetic small 9 ~( i- e, `5 }7 Z
interfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-8 O$ Z, G% {5 z, q
leagues in 2001, has further boosted the development of novel therapeutics and experi-
5 g, f* K7 U9 z5 }$ Z1 _1 o7 X( xmental tools based on RNAi technology.
- s/ ^- S# Q g3 ^& M9 B; {6 RViruses and RNAi share an intricate relationship at many levels. Early work in plants
* B# B- x1 n# v, I' V6 W& }! w2 Windicated that viruses can be both inducers and targets of RNA-based post-transcriptional
$ D4 O* E( |- u1 c! ogene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as
% w6 p( g; D; J+ i5 z* \an antiviral defense mechanism is now well-established in plants and other organisms,
" p6 z2 z( }% |; E, K3 cincluding insects. In vertebrates, viruses also interact with a related RNA silencing mecha-
+ Z% Q8 C) B" g5 e0 C3 x( ?* a2 n+ ynism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set 8 \' M( t3 o. U( M7 _6 Z* L* f9 g
of miRNAs, by which they regulate viral or host gene expression and modify, for example, ; L3 X. J" k$ \/ O8 V, m5 H6 ]7 ?* W
the transition from latent to lytic infection and the recognition of infected cells by the host 7 i3 {. F5 M. O6 F9 T
immune system. Furthermore, cellular miRNAs likely regulate expression of many genes 1 u; {; u& L% K* B) @
that are important for virus biology, but they have been suggested to directly target viral " ^3 M: B6 I" k) N
RNA as well.# W- c1 R$ t0 w4 n- o# @
The therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is 6 m/ [7 X& U# W" z! j' ^
now clear that replication of many, if not all, mammalian viruses can be suppressed by 7 Y3 v% N0 l; r( v) f# C
RNAi in cell culture. While these results have raised considerable optimism about the
) m+ O; F* r' S) qpotential of RNAi-based drugs, important hurdles remain, including issues related to the
0 m" x' Y3 x3 t' w% S7 S' J Ldelivery and stability of siRNAs and the risk of viral escape.0 e$ v" F8 q9 b8 |
From this brief overview it will be apparent that a great — and increasing — number . D$ v" ]+ K/ }; q
of tools and techniques are available for those interested in the interface of viruses and
) L. Q( T" v" ? \( TRNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-- m$ |$ ?+ j+ I. q
cols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for 6 j/ T- v) Y2 \0 Z! u- |' F$ u
the development and optimization of RNAi-based antiviral drugs. As RNAi is a central / Z9 m: x0 X O8 _
regulatory mechanism in the cell, the methods in this volume can also be applied out of # E H' L! o$ o+ U; u
the context of a virus infection. In the established tradition of the Methods in Molecular
( k7 c& i8 |5 X2 s6 UBiology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-% u$ G4 A$ l# Q7 Z6 W/ T' P! @2 @
by-step protocols and extra tools and tricks that should be useful to those new to the feld # z6 s; A) _$ C
and experienced scientists alike.
) v+ E" x' P& X* tThis volume consists of fve parts. Part 1 reviews important basic concepts in the feld # w7 k& |- @% j z, D; T* b( L
of antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of $ {8 b& P& e4 f- D) j, l: |: X# e
small silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-0 V* z7 Y% F! U% w7 R, M' D
ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
# t! T2 E. z8 G, F/ I, texpression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide
3 T5 z; T# i; v$ V4 ]RNAi approaches for the identifcation of factors involved in virus replication, which may ! D2 U! h) ~' w9 {
represent novel targets for antiviral therapy.
5 f# Y4 ?6 B2 W; [$ d3 bI am grateful to all authors for providing their outstanding contributions and to John * p$ U/ h- z8 F+ d }( v% D# p
Walker for guidance while editing this volume. I thank members of my lab, especially $ H. B7 m! [( l5 s
Walter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-, z9 | B8 r! {7 _8 ^* j
sions. I am thankful to Raul Andino for having been a great mentor and for introducing 9 {$ E+ {$ W$ Y' [
me to this exciting feld of research. Finally, I would like to apologize for doing little jus-, g w! t! V6 d. ~, [1 |
tice to the seminal work in plants; space limitations forced me to focus this volume on the & o+ k7 [% r# ^0 U+ ^2 q; P
animal system.0 k4 Y9 K* ]7 \; l( I+ |2 q
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
