Key Points
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A new method for elucidating gene function in mammals was discovered in 2000; it uses small double-stranded RNAs to interfere with gene expression.
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An evolutionarily conserved pathway uses these small interfering RNAs (siRNAs) to degrade mRNAs before translation.
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siRNAs are easily synthesized and used to silence genes in cell culture. Investigators are seizing the opportunity to use siRNA technology to silence their favourite genes. Recently, the successful use of siRNA in whole mice was reported.
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It has recently been shown that siRNAs can be effective antiviral agents in cell culture, raising hopes that siRNAs will represent the next generation of antiviral therapeutics.
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siRNAs can be expressed from DNA constructs and silencing cell lines can be made.
Abstract
Among the 3 billion base pairs of the human genome, there are ∼30,000–40,000 protein-coding genes, but the function of at least half of them remains unknown. A new tool — short interfering RNAs (siRNAs) — has now been developed for systematically deciphering the functions and interactions of these thousands of genes. siRNAs are an intermediate of RNA interference, the process by which double-stranded RNA silences homologous genes. Although the use of siRNAs to silence genes in vertebrate cells was only reported a year ago, the emerging literature indicates that most vertebrate genes can be studied with this technology.
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Acknowledgements
The authors thank P. Zanmore, B. Cullen, A. Pasquirelli and the Sharp lab for critical reading of the manuscript. Our apologies to those whose work which could not be cited due to space constraints. NCI grant to P.A.S. and partially by NCI Cancer centre support. M.T.M is a fellow of the Cancer Research Institute.
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Glossary
- FORWARD GENETICS
-
A genetic analysis that proceeds from phenotype to genotype by positional cloning or candidate-gene analysis.
- REVERSE GENETICS
-
A genetic analysis that proceeds from genotype to phenotype by gene-manipulation techniques, such as homologous recombination in embryonic stem cells.
- RIBOZYME
-
An RNA molecule with catalytic activity.
- POLYSOME
-
A functional unit of protein synthesis that consists of several ribosomes that are attached along the length of a single molecule of RNA.
- DYAD SYMMETRY
-
A sequence is said to be dyad symmetric (or pallindromic) when it is symmetric about a central axis.
- PRIMARY CELL
-
A cell that is taken from a tissue source and whose progeny are grown in culture, before subdivision and transfer to a subculture.
- CONFLUENCE
-
The degree at which a monolayer of tissue culture cells occupies the growth dish.
- DEAD BOX
-
A highly conserved motif in a family of putative RNA helicases; it takes its name from the single letter code for its amino-acid sequence.
- RESPIRATORY SYNCYTIAL VIRUS
-
A virus that is responsible for many respiratory tract infections, including the common cold.
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McManus, M., Sharp, P. Gene silencing in mammals by small interfering RNAs. Nat Rev Genet 3, 737–747 (2002). https://doi.org/10.1038/nrg908
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DOI: https://doi.org/10.1038/nrg908
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