siRNA as a tool for streamlining functional genomic studies

Abstract RNA interference (RNAi) has the potential to accelerate greatly the pace of discovery biology. The active RNAi intermediate is the small interfering RNA (siRNA), a discrete nucleic acid duplex that can be generated by several methods and used to directly silence gene expression. The choice of methods employed depends largely on the research or therapeutic objective. In most cases, rational design offers several advantages over random design, including greater predictability of function, higher silencing potency and longer duration of suppression. Of the production methods, chemical synthesis provides the fastest production capability, the highest purity and the easiest scalability for high-throughput strategies. Effective coupling of several methods gives the greatest potential for the application of RNAi across functional genomic and target validation studies. Furthermore, the coupling of RNAi with cellular profiling technologies will provide opportunities to streamline drug discovery significantly.

[1]  D. Baltimore,et al.  Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5 , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Stacy L DeRuiter,et al.  RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Sean D. Taverna,et al.  Methylation of Histone H3 at Lysine 9 Targets Programmed DNA Elimination in Tetrahymena , 2002, Cell.

[4]  A. Pasquinelli,et al.  Genes and Mechanisms Related to RNA Interference Regulate Expression of the Small Temporal RNAs that Control C. elegans Developmental Timing , 2001, Cell.

[5]  Michael T. McManus,et al.  Gene silencing using micro-RNA designed hairpins. , 2002, RNA.

[6]  M. Caruthers,et al.  Novel RNA Synthesis Method Using 5‘-O-Silyl-2‘-O-orthoester Protecting Groups , 1998 .

[7]  C. Napoli,et al.  Introduction of a Chimeric Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans. , 1990, The Plant cell.

[8]  J. Rossi,et al.  Functional siRNA expression from transfected PCR products. , 2002, RNA.

[9]  R. Bernards,et al.  A System for Stable Expression of Short Interfering RNAs in Mammalian Cells , 2002, Science.

[10]  S. Scaringe,et al.  RNA oligonucleotide synthesis via 5'-silyl-2'-orthoester chemistry. , 2001, Methods.

[11]  S. Hammond,et al.  An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells , 2000, Nature.

[12]  T. Fujisawa,et al.  Analysis of a piwi-Related Gene Implicates Small RNAs in Genome Rearrangement in Tetrahymena , 2002, Cell.

[13]  A. Pasquinelli,et al.  A Cellular Function for the RNA-Interference Enzyme Dicer in the Maturation of the let-7 Small Temporal RNA , 2001, Science.

[14]  G. Hannon RNA interference : RNA , 2002 .

[15]  Michael T. McManus,et al.  Small Interfering RNA-Mediated Gene Silencing in T Lymphocytes1 , 2002, The Journal of Immunology.

[16]  R. Cedergren,et al.  The automated chemical synthesis of long oligoribuncleotides using 2'-O-silylated ribonucleoside 3'-O-phosphoramidites on a controlled-pore glass support: synthesis of a 43-nucleotide sequence similar to the 3'-half molecule of an Escherichia coli formylmethionine tRNA , 1987 .

[17]  David L. Lewis,et al.  Efficient delivery of siRNA for inhibition of gene expression in postnatal mice , 2002, Nature Genetics.

[18]  G. Macino,et al.  Suppression of gene expression by homologous transgenes , 2004, Antonie van Leeuwenhoek.

[19]  W. Forrester,et al.  A DNA vector-based RNAi technology to suppress gene expression in mammalian cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Douglas S. Conklin,et al.  Gene expression: RNA interference in adult mice , 2002, Nature.

[21]  G. Macino,et al.  Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences , 1992, Molecular microbiology.

[22]  Ali Ehsani,et al.  Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells , 2002, Nature Biotechnology.

[23]  S. Scaringe Advanced 5'-silyl-2'-orthoester approach to RNA oligonucleotide synthesis. , 2000, Methods in enzymology.

[24]  Ronald H. A. Plasterk,et al.  RNA Silencing: The Genome's Immune System , 2002, Science.

[25]  F. Buchholz,et al.  Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Andrew Fire,et al.  The rde-1 Gene, RNA Interference, and Transposon Silencing in C. elegans , 1999, Cell.

[27]  R. Medzhitov,et al.  Retroviral delivery of small interfering RNA into primary cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[28]  B. Bass,et al.  A Role for the RNase III Enzyme DCR-1 in RNA Interference and Germ Line Development in Caenorhabditis elegans , 2001, Science.

[29]  Martin Tabler,et al.  Short 5′-phosphorylated double-stranded RNAs induce RNA interference in Drosophila , 2001, Current Biology.

[30]  D. Engelke,et al.  Effective expression of small interfering RNA in human cells , 2002, Nature Biotechnology.

[31]  M. Caruthers,et al.  Gene synthesis machines: DNA chemistry and its uses. , 1985, Science.

[32]  P. Silver,et al.  Retrovirus-delivered siRNA , 2002, BMC biotechnology.

[33]  A. Caudy,et al.  Role for a bidentate ribonuclease in the initiation step of RNA interference , 2001 .

[34]  S. Pitsch,et al.  Synthesis and pairing properties of oligoribonucleotide analogues containing a metal-binding site attached to beta-D-allofuranosyl cytosine. , 1998, Nucleic acids research.

[35]  Titia Sijen,et al.  On the Role of RNA Amplification in dsRNA-Triggered Gene Silencing , 2001, Cell.

[36]  A. Fire,et al.  Functional anatomy of a dsRNA trigger: differential requirement for the two trigger strands in RNA interference. , 2000, Molecular cell.