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本帖最后由 细胞海洋 于 2011-6-25 11:08 编辑 5 ?* i+ N. p) z1 H
! }: d& O3 y) D; e* x) BPreface
1 g4 t% R) {* z) XLittle over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that % a5 ~+ h7 e' h3 K% `
double-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode
4 w0 m3 S5 |! W! A) H' ~/ CCaenorhabditis elegans (RNA interference, RNAi). This work converged with research in ) p# u9 P% D5 f
plants, in which related RNA-based silencing processes were known to exist. Ever since,
; |$ b' |4 N+ aresearch in the feld has progressed at an astonishing rate, resulting in our appreciation of
$ X7 ? R2 `0 \& O: O p/ Vsmall silencing RNAs as central regulators of gene expression, as guards of genome integ-9 }0 ^$ S& { z4 Y& {
rity, and as essential mediators of antiviral defense. The discovery that synthetic small
% A" Y, U) T8 u/ _ a/ ?2 t) Ginterfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-4 T$ h; V0 a8 Y& ]- [4 [/ F2 n2 P
leagues in 2001, has further boosted the development of novel therapeutics and experi-0 O9 R7 \# I, [$ f
mental tools based on RNAi technology.
7 r; w' y* H3 y" TViruses and RNAi share an intricate relationship at many levels. Early work in plants 2 a i/ R9 ~% M# F
indicated that viruses can be both inducers and targets of RNA-based post-transcriptional
" v! }% ~8 C% U$ pgene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as - d& D4 `5 o$ ~" M3 w3 E
an antiviral defense mechanism is now well-established in plants and other organisms,
6 v( [2 f. ]) ^, K4 B: dincluding insects. In vertebrates, viruses also interact with a related RNA silencing mecha-4 C, o$ p$ g! W* }7 | W
nism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set % p- ]# o) J6 \# E, J% \
of miRNAs, by which they regulate viral or host gene expression and modify, for example, . c+ j1 f2 U# P$ A7 j: D
the transition from latent to lytic infection and the recognition of infected cells by the host
$ b/ L7 \$ @" ` Qimmune system. Furthermore, cellular miRNAs likely regulate expression of many genes 7 X" k$ o( R, `% p6 S
that are important for virus biology, but they have been suggested to directly target viral & d& F [ ^$ V% j3 H$ L
RNA as well.
3 f5 h5 C4 k/ V7 {. y4 k% v9 GThe therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is ' v# V- Q( _& z9 e0 d2 _/ y
now clear that replication of many, if not all, mammalian viruses can be suppressed by
1 } Z$ s# c# ~RNAi in cell culture. While these results have raised considerable optimism about the
" r9 W1 C! H: Jpotential of RNAi-based drugs, important hurdles remain, including issues related to the 4 u' A% i) Q) T4 [8 ^
delivery and stability of siRNAs and the risk of viral escape.# P! e# h9 b9 e/ C% o/ a
From this brief overview it will be apparent that a great — and increasing — number
1 L; L4 n7 b0 J6 o1 zof tools and techniques are available for those interested in the interface of viruses and
" f" W0 v: b, w- U; aRNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-* s" N0 D) X& }) X# j. O0 x
cols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for $ j- X: E' d7 }7 G% S; A
the development and optimization of RNAi-based antiviral drugs. As RNAi is a central " q" P9 n) h$ I8 O
regulatory mechanism in the cell, the methods in this volume can also be applied out of
/ B* M' Q! ^. T7 ]the context of a virus infection. In the established tradition of the Methods in Molecular
) c4 v! a3 |/ `7 P5 kBiology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-+ G" u$ p) T! S# M4 q: k `$ N0 v! G
by-step protocols and extra tools and tricks that should be useful to those new to the feld ; V. l4 e4 J2 y
and experienced scientists alike.# i3 `, n& [. @8 c) [
This volume consists of fve parts. Part 1 reviews important basic concepts in the feld
& j" ^, d) [" q& d1 q0 `3 Vof antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of 9 w/ W0 L. ]; ?9 S
small silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-4 e P: I6 V& |" O& P( ^
ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
0 k; o/ { T- A! h$ hexpression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide " n4 V7 F& x- ~2 }2 S4 v" {
RNAi approaches for the identifcation of factors involved in virus replication, which may & P1 F) w# h3 [8 T" q: O
represent novel targets for antiviral therapy.# C% N5 q8 E. j, w) k! K
I am grateful to all authors for providing their outstanding contributions and to John 8 [# V/ T- Z g. x5 G/ m3 J4 P
Walker for guidance while editing this volume. I thank members of my lab, especially
. e {8 T6 r+ `: u+ I/ \Walter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-
1 F$ y) }" s* h U. rsions. I am thankful to Raul Andino for having been a great mentor and for introducing 2 b/ U& ?* t; Y. }
me to this exciting feld of research. Finally, I would like to apologize for doing little jus-$ [" K! @ W# b# n! @
tice to the seminal work in plants; space limitations forced me to focus this volume on the 0 \# r, W, }; S. z* D4 `
animal system.& Y* C2 r! y, b4 P
Nijmegen, The Netherlands Ronald P. van Rij |
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发表于 2011-6-25 10:44
