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[实验技术类] PDF电子书:Antiviral RNAi - Concepts, Methods, and Applications 2011     [复制链接]

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发表于 2011-6-25 10:44 |只看该作者 |正序浏览 |打印
本帖最后由 细胞海洋 于 2011-6-25 11:08 编辑 ' }# Y4 z" ~. R" x. {5 N' J1 }
6 v  g% e! D4 e: m' P% w5 u9 i1 T! j
Preface
$ t6 Y( W6 `6 ^+ \+ u# wLittle over a decade ago, Andrew Fire, Craig Mello, and colleagues demonstrated  that 6 Y* ]: o& s) c; X5 V
double-stranded  (ds)RNA  induces  sequence-specifc  gene  silencing  in  the  nematode
5 W6 e5 b  R/ q" W( lCaenorhabditis elegans (RNA interference, RNAi). This work converged with research in
& z- K+ G+ Z5 z. M3 d  j! A+ aplants, in which related RNA-based silencing processes were known to exist. Ever since, 9 N+ A6 a7 x1 c. N
research in the feld has progressed at an astonishing rate, resulting in our appreciation of
' ]* M1 g3 ?' i" L5 w& Vsmall silencing RNAs as central regulators of gene expression, as guards of genome integ-
/ j. {% i/ K* Urity,  and  as  essential mediators  of  antiviral  defense. The  discovery  that  synthetic  small 1 d* [9 D6 q+ X( Q
interfering RNA (siRNA) induces gene silencing in mammals, by Thomas Tuschl and col-' C) `+ v, P+ b
leagues in 2001, has further boosted the development of novel therapeutics and experi-
& U, K- O# }) I, V" T" m/ hmental tools based on RNAi technology.( F, r9 i9 T, e/ P. ]4 w, @6 o
Viruses and RNAi share an intricate relationship at many levels. Early work in plants
1 S0 ~3 C8 Y4 Y0 `) Vindicated that viruses can be both inducers and targets of RNA-based post-transcriptional
) ^& @5 t0 t6 |' \- qgene silencing (which we now know as RNAi or RNA silencing). The concept of RNAi as " Z% p1 D6 j  ^2 K" m1 L( w" h
an  antiviral  defense mechanism  is  now well-established  in  plants  and  other  organisms,
3 w; I# _2 N8 j4 V3 M1 r, t+ cincluding insects. In vertebrates, viruses also interact with a related RNA silencing mecha-* ~* ~# J$ ~: `2 G2 E
nism, the microRNA (miRNA) pathway. Many nuclear DNA viruses encode their own set
/ K1 X: b0 g' p0 E9 m0 l% Mof miRNAs, by which they regulate viral or host gene expression and modify, for example,
) {: d5 l0 L4 c6 H3 t6 Wthe transition from latent to lytic infection and the recognition of infected cells by the host
4 K' ^3 @5 ]& P" ?immune system. Furthermore, cellular miRNAs likely regulate expression of many genes % W' A* Z8 Z4 w
that are important for virus biology, but they have been suggested to directly target viral
8 q; g# k' v- [% O& ?RNA as well.4 [2 T5 ~8 t+ \* o3 ~+ W
The therapeutic potential of RNAi-based antiviral drugs was recognized early on. It is ; s5 b; Y/ A' l, j1 x
now clear  that  replication of many,  if not all, mammalian viruses can be  suppressed by
5 }' V' u$ ^$ @8 z6 T5 p0 H5 xRNAi  in  cell  culture. While  these  results have  raised  considerable optimism  about  the , h7 s8 Y. n1 o7 p  R& o7 q- Z
potential of RNAi-based drugs, important hurdles remain, including issues related to the
5 k2 J5 ]; R; ]2 u& A2 Jdelivery and stability of siRNAs and the risk of viral escape.
- P: G: Z* W6 @3 \- Y) zFrom this brief overview it will be apparent that a great — and increasing — number 5 `+ ~; a0 e! t+ Z6 V
of  tools and  techniques are available  for  those  interested  in  the  interface of viruses and
3 N' m' D8 O( K$ L3 ORNAi. Antiviral RNAi: concepts, methods, and applications provides a collection of proto-
+ ~! G, a" D( Y: d% T9 a' Q' Ycols for the analysis of natural antiviral RNAi responses and viral miRNAs, as well as for * D- L. ?4 U5 a$ z
the development and optimization of RNAi-based antiviral drugs. As RNAi  is a central
7 P2 k  M6 E: H* }regulatory mechanism in the cell, the methods in this volume can also be applied out of 4 k+ u) z! A- D
the context of a virus infection. In the established tradition of the Methods in Molecular . @$ I$ C+ y9 j0 F
Biology series, Antiviral RNAi: concepts, methods, and applications provides detailed step-) ^4 I2 a* p  @2 C
by-step protocols and extra tools and tricks that should be useful to those new to the feld
' b( T' U7 K& j2 hand experienced scientists alike.
' C1 |' `3 A: y0 _This volume consists of fve parts. Part 1 reviews important basic concepts in the feld
  d: l- W2 {2 L) l) u+ i# Oof antiviral RNAi. Part 2 provides experimental and bio-informatic tools for the analysis of 2 ]+ y% {; Y" G# F' e2 H
small silencing RNAs. Part 3 covers methods to biochemically dissect RNAi-based antivi-; @& m# k- g  f, X3 ?
ral defense and viral counter-defense mechanisms. Part 4 describes methods for the design,
, R' Q1 x  ]: T! ~( n' E$ Aexpression,  and delivery of  therapeutic  antiviral  siRNAs. Part 5  presents  genome-wide ' O, {- n8 F( }$ y7 V
RNAi approaches for the identifcation of factors involved in virus replication, which may
$ u/ R6 L5 d; T5 r4 P& S5 S5 X; ^8 |represent novel targets for antiviral therapy.' T# Q+ L' R( w& X7 L# W
I am grateful to all authors for providing their outstanding contributions and to John
  l, D; D3 V( aWalker  for guidance while  editing  this  volume.  I  thank members of my  lab,  especially
* ]& t7 R9 @" y$ @Walter Bronkhorst, Koen van Cleef, Marius van den Beek, and Joël van Mierlo, for discus-
2 \4 ~5 ]% @$ C$ Q& Xsions. I am thankful to Raul Andino for having been a great mentor and for introducing ) f3 f9 P( K9 q3 d' x
me to this exciting feld of research. Finally, I would like to apologize for doing little jus-: G& a7 K: [7 }/ K+ f- c' |
tice to the seminal work in plants; space limitations forced me to focus this volume on the
/ I! h6 r% e' u, r8 e) z4 d  Tanimal system.( A- _$ `' }% \* q: m
Nijmegen, The Netherlands  Ronald P. van Rij

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我在努力中  

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干细胞之家微信公众号
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感谢党和人民的关爱~~~  

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风物长宜放眼量  

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原来这样也可以  
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