Article Information

• PDF (322 kb)
• Full Text with Thumbnail Figures
• Full Text with Large Figures
• Cited by in Scopus (77)

PubMed

• Articles by Konrad Huppi
• Articles by Natasha J. Caplen

Related Articles

• RNA interference: the new somatic cell genetics?

  RNA interference: the new somatic cell genetics? Cancer Cell, Volume 2, Issue 1, 1 July 2002, Pages 17-23 Patrick J Paddison and Gregory J Hannon Summary RNAi is evolving into a powerful tool for manipulating gene expression in mammalian cells with potential utility for investigating gene function, for high-throughput, function-based genetic screens and potentially for development as a therapeutic tool. Summary | Full Text | PDF (276 kb)

• The silent treatment: RNAi as a defense against virus infect...

  The silent treatment: RNAi as a defense against virus infection in mammals Trends in Biotechnology, Volume 24, Issue 4, 1 April 2006, Pages 186-193 Ronald P. van Rij and Raul Andino Abstract RNA interference (RNAi) is a mechanism for sequence-specific gene silencing guided by double-stranded RNA. In plants and insects it is well established that RNAi is instrumental in the response to viral infections; whether RNAi has a similar function in mammals is under intense investigation. Recent studies to address this question have identified some unanticipated interactions between the RNAi machinery and mammalian viruses. Furthermore, introduction of virus-specific small interfering RNAs (siRNAs) into cells, thus programming the RNAi machinery to target viruses, is an effective therapeutic approach to inhibit virus replication and in animal models. Although several issues remain to be addressed, such as delivery and viral escape, these findings hold tremendous potential for the development of RNAi-based antiviral therapeutics. Abstract | Full Text | PDF (193 kb)

• Small non-coding RNAs as magic bullets

  Small non-coding RNAs as magic bullets Trends in Biochemical Sciences, Volume 30, Issue 8, 1 August 2005, Pages 445-452 Fritz Eckstein Abstract RNA interference (RNAi) – inhibition of gene expression by small, non-coding RNAs [small interfering RNAs (siRNAs) or microRNAs (miRNAs)] – has changed our view of regulation of expression dramatically. The application of siRNAs for both functional analysis of genes and medication raises several questions. These include the design of the double-stranded oligonucleotides, their preparation and introduction into cells or animals either as chemically synthesized entities or as transcripts from a suitable vector. Delivery of the oligonucleotides, choice of vector, chemical modification to stabilize against nucleases and avoidance of side effects (e.g. stimulation of interferons) are major challenges. Work to identify the multiple targets of miRNAs is still in its infancy, and a clear distinction between siRNAs and miRNAs is difficult in some instances. Moreover, transcriptional silencing by RNAi is poorly understood; it is evident that the siRNA machinery is involved but the details await clarification. Given the multitude of interactions of the small non-coding RNAs revealed so far, we should be prepared to encounter, as yet, undiscovered interactions and mechanisms. Abstract | Full Text | PDF (155 kb)

• …more

Defining and Assaying RNAi in Mammalian Cells

Konrad Huppi, Scott E. Martin and Natasha J. Caplen*^,  

Gene Silencing Section, Office of Science and Technology Partnerships, Office of the Director, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Building 37, 3128B, 37 Convent Drive, Bethesda, MD 20892 USA

Correspondence: Natasha J. Caplen, 301-451-1844 (phone), 301-402-1031 (fax)

Summary

The investigation of protein function through the inhibition of activity has been critical to our understanding of many normal and abnormal biological processes. Until recently, functional inhibition in biological systems has been induced using a variety of approaches including small molecule antagonists, antibodies, aptamers, ribozymes, antisense oligonucleotides or transcripts, morpholinos, dominant-negative mutants, and knockout transgenic animals. Although all of these approaches have made substantial advances in our understanding of the function of many proteins, a lack of specificity or restricted applicability has limited their utility. Recently, exploitation of the naturally occurring posttranscriptional gene silencing mechanism triggered by double-stranded RNA (dsRNA), termed RNA interference (RNAi), has gained much favor as an alternative means for analyzing gene function. Aspects of the basic biology of RNAi, its application as a functional genomics tool, and its potential as a therapeutic approach have been extensively reviewed (Hannon and Rossi, 2004; Meister and Tuschl, 2004); however, there has been only limited discussion as to how to design and validate an individual RNAi effector molecule and how to interpret RNAi data overall, particularly with reference to experimentation in mammalian cells. This perspective will aim to consider some of the issues encountered when conducting and interpreting RNAi experiments in mammalian cells.