Journal ArticleDOI
U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells
Reads0
Chats0
TLDR
A vector-based siRNA expression system that can induce RNAi in mammalian cells is reported, which might allow therapeutic applications by means of vector-mediated RNAi and facilitate a wide range of functional analysis of mammalian genes.Abstract:
The first evidence for gene disruption by double-stranded RNA (dsRNA) came from careful analysis in Caenorhabditis elegans. This phenomenon, called RNA interference (RNAi), was observed subsequently in various organisms, including plants, nematodes, Drosophila, and protozoans. Very recently, it has been reported that in mammalian cells, 21- or 22-nucleotide (nt) RNAs with 2-nt 3' overhangs (small inhibitory RNAs, siRNAs) exhibit an RNAi effect. This is because siRNAs are not recognized by the well-characterized host defense system against viral infections, involving dsRNA-dependent inhibition of protein synthesis. However, the current method for introducing synthetic siRNA into cells by lipofection restricts the range of applications of RNAi as a result of the low transfection efficiencies in some cell types and/or short-term persistence of silencing effects. Here, we report a vector-based siRNA expression system that can induce RNAi in mammalian cells. This technical advance for silencing gene expression not only facilitates a wide range of functional analysis of mammalian genes but might also allow therapeutic applications by means of vector-mediated RNAi.read more
Citations
More filters
Journal ArticleDOI
RNA-Guided Human Genome Engineering via Cas9
Prashant Mali,Luhan Yang,Kevin M. Esvelt,John Aach,Marc Güell,James E. DiCarlo,Julie E. Norville,George M. Church,George M. Church +8 more
TL;DR: The type II bacterial CRISPR system is engineer to function with custom guide RNA (gRNA) in human cells to establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.
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
Cloning of adiponectin receptors that mediate antidiabetic metabolic effects
Toshimasa Yamauchi,Junji Kamon,Yusuke Ito,Atsushi Tsuchida,Takehiko Yokomizo,Takehiko Yokomizo,Shunbun Kita,Takuya Sugiyama,Makoto Miyagishi,Makoto Miyagishi,Kazuo Hara,Masaki Tsunoda,Koji Murakami,Toshiaki Ohteki,S. Uchida,Sato Takekawa,Hironori Waki,Nelson H. Tsuno,Yoichi Shibata,Yasuo Terauchi,Philippe Froguel,Kazuyuki Tobe,Shigeo Koyasu,Kazunari Taira,Kazunari Taira,Toshio Kitamura,Takao Shimizu,Takao Shimizu,Ryozo Nagai,Takashi Kadowaki +29 more
TL;DR: The cloning of complementary DNAs encoding adiponectin receptors 1 and 2 by expression cloning supports the conclusion that they serve as receptors for globular and full-length adiponECTin, and that they mediate increased AMP kinase and PPAR-α ligand activities, as well as fatty-acid oxidation and glucose uptake by adiponectionin.
Journal ArticleDOI
A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference
Douglas A. Rubinson,Christopher P. Dillon,Adam V. Kwiatkowski,Claudia Sievers,Claudia Sievers,Lili Yang,Johnny E. Kopinja,Dina L Rooney,Mingdi Zhang,Melanie Ihrig,Michael T. McManus,Frank B. Gertler,Martin L. Scott,Luk Van Parijs +13 more
TL;DR: It is shown that lentivirus-delivered shRNAs are capable of specific, highly stable and functional silencing of gene expression in a variety of cell types and also in transgenic mice.
Journal ArticleDOI
Gene silencing in mammals by small interfering RNAs
TL;DR: This work has shown that the use of siRNAs to silence genes in vertebrate cells was only reported a year ago, and the emerging literature indicates that most vertebrate genes can be studied with this technology.
References
More filters
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
TL;DR: 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.
Journal ArticleDOI
Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells
TL;DR: 21-nucleotide siRNA duplexes provide a new tool for studying gene function in mammalian cells and may eventually be used as gene-specific therapeutics.
Journal ArticleDOI
Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate.
TL;DR: Duplexes of 21–23 nucleotide RNAs are the sequence‐specific mediators of RNA interference and post‐transcriptional gene silencing and mismatches in the centre of the siRNA duplex prevent target RNA cleavage, providing a rational basis for the design of siRNAs in future gene targeting experiments.
Journal ArticleDOI
Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems.
TL;DR: Synthetic siRNAs can induce gene-specific inhibition of expression in Caenorhabditis elegans and in cell lines from humans and mice, and seem to avoid the well documented nonspecific effects triggered by longer double-stranded RNAs in mammalian cells.
Journal ArticleDOI
Wnt Signaling in Oncogenesis and Embryogenesis--a Look Outside the Nucleus
Mark Peifer,Paul Polakis +1 more
TL;DR: Two areas of rapid advance are focused on: the machinery that regulates the stability of the key signal transducer, β-catenin, and the effect of Wnt signaling on cellular targets outside the nucleus, the actin and microtubule cytoskeletons.