Journal ArticleDOI
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Andrew Fire,SiQun Xu,Mary K. Montgomery,Steven A. Kostas,Steven A. Kostas,Samuel E. Driver,Craig C. Mello +6 more
TLDR
To their surprise, it was found that double-stranded RNA was substantially more effective at producing interference than was either strand individually, arguing against stochiometric interference with endogenous mRNA and suggesting that there could be a catalytic or amplification component in the interference process.Abstract:
Experimental introduction of RNA into cells can be used in certain biological systems to interfere with the function of an endogenous gene Such effects have been proposed to result from a simple antisense mechanism that depends on hybridization between the injected RNA and endogenous messenger RNA transcripts RNA interference has been used in the nematode Caenorhabditis elegans to manipulate gene expression Here we investigate the requirements for structure and delivery of the interfering RNA To our surprise, we found that double-stranded RNA was substantially more effective at producing interference than was either strand individually After injection into adult animals, purified single strands had at most a modest effect, whereas double-stranded mixtures caused potent and specific interference The effects of this interference were evident in both the injected animals and their progeny Only a few molecules of injected double-stranded RNA were required per affected cell, arguing against stochiometric interference with endogenous mRNA and suggesting that there could be a catalytic or amplification component in the interference processread more
Citations
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MicroRNAs: Genomics, Biogenesis, Mechanism, and Function
TL;DR: Although they escaped notice until relatively recently, miRNAs comprise one of the more abundant classes of gene regulatory molecules in multicellular organisms and likely influence the output of many protein-coding genes.
Journal ArticleDOI
RNA interference is mediated by 21- and 22-nucleotide RNAs
TL;DR: In this article, the authors demonstrate that 21 and 22-nt RNA fragments are the sequence-specific mediators of RNA interference in a Drosophila in vitro system, and provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the siRNA-protein complex.
Journal ArticleDOI
Systematic functional analysis of the Caenorhabditis elegans genome using RNAi
Ravi S. Kamath,Andrew G. Fraser,Andrew G. Fraser,Yan Dong,Gino B. Poulin,Richard Durbin,Monica Gotta,Alexander Kanapin,Nathalie Le Bot,Sergio Moreno,Sergio Moreno,Marc Sohrmann,David P. Welchman,Peder Zipperlen,Julie Ahringer +14 more
TL;DR: It is found that genes of similar functions are clustered in distinct, multi-megabase regions of individual chromosomes; genes in these regions tend to share transcriptional profiles.
Journal ArticleDOI
A species of small antisense RNA in posttranscriptional gene silencing in plants.
TL;DR: The 25-nucleotide antisense RNA detected in transgene-induced PTGS is likely synthesized from an RNA template and may represent the specificity determinant of PTGS.
Journal ArticleDOI
Exportin-5 mediates the nuclear export of pre-microRNAs and short hairpin RNAs
TL;DR: It is demonstrated that human pre-miRNA nuclear export, and miRNA function, are dependent on Exportin-5, an additional cellular cofactor required for miRNA biogenesis and function.
References
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Journal ArticleDOI
The genetics of caenorhabditis elegans
TL;DR: In this paper, the authors describe methods for the isolation, complementation and mapping of mutants of Caenorhabditis elegans, a small free-living nematode worm.
Journal ArticleDOI
Green fluorescent protein as a marker for gene expression
TL;DR: A complementary DNA for the Aequorea victoria green fluorescent protein produces a fluorescent product when expressed in prokaryotic or eukaryotic cells, which can be used to monitor gene expression and protein localization in living organisms.
Journal Article
The genetics of Caenorhabditis elegans.
TL;DR: Estimates of the induced mutation frequency of both the visible mutants and X chromosome lethals suggests that, just as in Drosophila, the genetic units in C. elegans are large.
Journal ArticleDOI
The embryonic cell lineage of the nematode Caenorhabditis elegans.
TL;DR: It is concluded that the cell lineage itself, complex as it is, plays an important role in determining cell fate and is demonstrated to demonstrate substantial cell autonomy in at least some sections of embryogenesis.
Journal ArticleDOI
par-1, a Gene Required for Establishing Polarity in C. Elegans Embryos, Encodes a Putative Ser/Thr Kinase That Is Asymmetrically Distributed
Su Guo,Kenneth J. Kemphues +1 more
TL;DR: It is reported here that par-1 encodes a putative Ser/Thr kinase with similarity to kinases from yeasts and mammals, suggesting that kinase activity is essential for par-2 function and in establishing embryonic polarity.