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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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Preface
! A, C" M5 X' Z2 A2 f, V' ZLittle over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that
) u; b* [1 a3 Q& M1 I7 Zdouble-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode
1 K1 }3 X5 G: Q% g' H" m7 _0 TCaenorhabditis elegans (RNA interference, RNAi). This work converged with research in ) q' P9 U" l) c& W
plants, in which related RNA-based silencing processes were known to exist. Ever since,
1 Z9 M& q( T9 C+ Lresearch in the feld has progressed at an astonishing rate, resulting in our appreciation of ( I0 l* u4 W/ ^, ~9 K: t3 i
small silencing RNAs as central regulators of gene expression, as guards of genome integ-
2 j" y6 ^5 z! krity, and as essential mediators of antiviral defense. The discovery that synthetic small 0 ]9 `9 G9 i/ W% w
interfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col- F5 v5 k; @# @! q4 \4 p
leagues in 2001, has further boosted the development of novel therapeutics and experi-
+ T& G9 M5 P3 S1 w# emental tools based on RNAi technology./ S$ N2 M8 g( `! b
Viruses and RNAi share an intricate relationship at many levels. Early work in plants
# t) Y" ^1 w- r( c8 T5 eindicated that viruses can be both inducers and targets of RNA-based post-transcriptional
; S( J! D7 T- K) ?gene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as 6 q7 @6 E; f+ V" _
an antiviral defense mechanism is now well-established in plants and other organisms,
' j3 O5 m2 M S6 g7 k# \including insects. In vertebrates, viruses also interact with a related RNA silencing mecha-
) K9 k9 q* ^* A9 }) A( ^' Xnism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set # k) m ]. j5 _9 l- W# P5 ~6 {
of miRNAs, by which they regulate viral or host gene expression and modify, for example, 2 F$ ^! f) w" _6 b
the transition from latent to lytic infection and the recognition of infected cells by the host
7 l& ?! p& X$ Z! pimmune system. Furthermore, cellular miRNAs likely regulate expression of many genes
$ A0 y& f* `- b& \& Othat are important for virus biology, but they have been suggested to directly target viral
! m+ Q# X" @( q+ ` O7 q, DRNA as well.3 J3 \9 Q0 [% U0 A' t
The therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is + E4 x: F. k7 a) b; F
now clear that replication of many, if not all, mammalian viruses can be suppressed by
2 E8 W& \9 O+ Y8 s- R( A0 H1 gRNAi in cell culture. While these results have raised considerable optimism about the
# q' O8 f; @6 q& vpotential of RNAi-based drugs, important hurdles remain, including issues related to the
0 Y" B- N4 d3 C2 P& i% Z$ n9 ~- ]delivery and stability of siRNAs and the risk of viral escape.; f0 f" K/ Y) y
From this brief overview it will be apparent that a great — and increasing — number
: E: @- Z) F- dof tools and techniques are available for those interested in the interface of viruses and 7 i& s, \- E2 v# J
RNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto- A7 a1 E; \% L# H7 w1 ^3 O
cols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for 7 q) d4 A z9 E7 s
the development and optimization of RNAi-based antiviral drugs. As RNAi is a central * r4 M8 ^8 b) m8 ]( Z, b
regulatory mechanism in the cell, the methods in this volume can also be applied out of
' n |8 B3 k$ Y! a* d! R! Jthe context of a virus infection. In the established tradition of the Methods in Molecular 2 P6 N+ P8 \1 B9 t% u
Biology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-- Q: f+ [, Q/ u% I1 p0 [
by-step protocols and extra tools and tricks that should be useful to those new to the feld
" M% J0 s7 E. S3 Q' Iand experienced scientists alike.5 ~; k0 E, u' k5 b; g& U
This volume consists of fve parts. Part 1 reviews important basic concepts in the feld 9 R, q1 J# v; H4 ^
of antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of ! ?6 D+ h2 f0 ^" i( |0 e6 L% L
small silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-
1 P1 Q( [5 w9 q' S" K, wral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
. f! I0 P4 m Q; f3 v8 F; w. D- x, e- Kexpression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide
2 F2 v# O# C4 q$ JRNAi approaches for the identifcation of factors involved in virus replication, which may
/ X- V% F" r Z/ y" ~" l) Y5 Q7 Urepresent novel targets for antiviral therapy.% a9 e5 G" h1 i: |) F
I am grateful to all authors for providing their outstanding contributions and to John
% Y: E( L u/ n$ qWalker for guidance while editing this volume. I thank members of my lab, especially 7 B: u- L. f( Z
Walter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-
. {% l6 M/ M/ lsions. I am thankful to Raul Andino for having been a great mentor and for introducing 6 J+ I3 N2 {/ o) ~
me to this exciting feld of research. Finally, I would like to apologize for doing little jus-3 c N ]6 W1 J8 r8 ~0 A& @% B& X# t
tice to the seminal work in plants; space limitations forced me to focus this volume on the 1 t' m+ x( _* W+ ]8 k0 ^& W
animal system.
3 X4 \& g; \/ \- v$ d$ @0 HNijmegen, 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
