|
 
- 积分
- 631
- 威望
- 631
- 包包
- 1519
|
本帖最后由 细胞海洋 于 2011-6-25 11:08 编辑
2 D; c( y8 W, D; U) B& M( {' t7 q% }; h' L, k9 g0 `9 o; I
Preface
- P" }; N" G2 X: ?5 R( K ?Little over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated that
. ]0 z T9 X% c1 I( | L4 Edouble-stranded (ds)RNA induces sequence-specifc gene silencing in the nematode 3 c1 d8 l/ U( c E. A$ U$ e
Caenorhabditis elegans (RNA interference, RNAi). This work converged with research in 9 {5 C' D) F; e$ k: O$ a0 P& ]
plants, in which related RNA-based silencing processes were known to exist. Ever since,
8 ~. N+ S1 a7 u L Bresearch in the feld has progressed at an astonishing rate, resulting in our appreciation of : _% K+ T- ` |6 ^0 L! n
small silencing RNAs as central regulators of gene expression, as guards of genome integ-8 N) Z& r, K" S; _- q8 Y: A
rity, and as essential mediators of antiviral defense. The discovery that synthetic small , _* i9 N$ ` }/ e: [) m
interfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-
7 [0 Y S: N0 Q) k- p2 S/ ~7 Z3 Bleagues in 2001, has further boosted the development of novel therapeutics and experi-, G: J6 m; R5 o6 I. [
mental tools based on RNAi technology.
0 J# [8 ~* h; f* kViruses and RNAi share an intricate relationship at many levels. Early work in plants 1 G0 D8 M0 N) r* w% F& J K
indicated that viruses can be both inducers and targets of RNA-based post-transcriptional ; J+ {. ]2 m/ d! K( X; m
gene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as
, i, C1 H9 S" H! v4 K/ C xan antiviral defense mechanism is now well-established in plants and other organisms,
9 J5 m0 C, e* j! ^& {: Y, E% oincluding insects. In vertebrates, viruses also interact with a related RNA silencing mecha-4 R% |( o+ Q9 `
nism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set
8 K* s+ t, w- W# x$ T8 d9 p6 X2 hof miRNAs, by which they regulate viral or host gene expression and modify, for example, ' Q' O- k. |( r: C- E Q
the transition from latent to lytic infection and the recognition of infected cells by the host 6 E7 Z" O% c3 a' k. ~& {- c5 c
immune system. Furthermore, cellular miRNAs likely regulate expression of many genes
( Y* _$ _1 J' v2 f3 u9 ^: _that are important for virus biology, but they have been suggested to directly target viral 7 Y L: T! i' n, c/ \
RNA as well.
. V, K5 B/ P) Z# lThe therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is
2 D& q4 T4 O @now clear that replication of many, if not all, mammalian viruses can be suppressed by " N7 T1 p/ b) z2 n
RNAi in cell culture. While these results have raised considerable optimism about the
/ _8 g% J" S$ u8 u, y3 Fpotential of RNAi-based drugs, important hurdles remain, including issues related to the
6 _. \, _' k+ F) F. ~delivery and stability of siRNAs and the risk of viral escape.
: T5 ^7 E ^; G5 iFrom this brief overview it will be apparent that a great — and increasing — number
. h' G" r/ P9 Xof tools and techniques are available for those interested in the interface of viruses and " j$ N U( `' @+ j0 [! H
RNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-
% v: G3 c. | N2 Ncols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for
s/ f% j. @) A- J0 d: Jthe development and optimization of RNAi-based antiviral drugs. As RNAi is a central
9 J4 U" P. c, ]$ l1 h+ Gregulatory mechanism in the cell, the methods in this volume can also be applied out of + S* `6 g# Z/ J$ D$ ^
the context of a virus infection. In the established tradition of the Methods in Molecular
* x. L; W' n/ f" P" OBiology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-
) c" D% y+ m8 B3 t5 ~by-step protocols and extra tools and tricks that should be useful to those new to the feld * u0 d: W* E/ A/ H7 w8 {: k3 `
and experienced scientists alike.5 W. n' C& `- D% i4 V7 A @
This volume consists of fve parts. Part 1 reviews important basic concepts in the feld
& g z& R K- b8 T6 tof antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of
6 e/ @2 O0 H& G& }# `' l; ksmall silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-# u. m) K- Q3 t4 Z3 E# f
ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
# o, d5 s' c( l% q# Yexpression, and delivery of therapeutic antiviral siRNAs. Part 5 presents genome-wide
2 _9 P+ E& I1 s4 l2 T0 hRNAi approaches for the identifcation of factors involved in virus replication, which may 0 ?% B) J- q7 O+ f! G
represent novel targets for antiviral therapy.6 w# a9 G" C+ {( c- s& k( D
I am grateful to all authors for providing their outstanding contributions and to John
' L6 a5 b9 X- Y, Y. A& u8 Y pWalker for guidance while editing this volume. I thank members of my lab, especially $ H6 M) R' W+ O9 ?8 p$ }
Walter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-
9 ] F! W- z3 {4 U4 xsions. I am thankful to Raul Andino for having been a great mentor and for introducing 0 d1 k: H8 l$ @+ O
me to this exciting feld of research. Finally, I would like to apologize for doing little jus-. \% L5 b- [5 y" B+ b8 ~
tice to the seminal work in plants; space limitations forced me to focus this volume on the * ~2 z' W K, ?
animal system.5 {; j( B5 D# d" k
Nijmegen, The Netherlands Ronald P. van Rij |
|
发表于 2011-6-25 10:44
发表于 2011-6-25 11:09
