Simple, robust strategies for generating DNA-directed RNA interference constructs.

@article{Rice2005SimpleRS,
  title={Simple, robust strategies for generating DNA-directed RNA interference constructs.},
  author={Robert R. Rice and A. Muirhead and Bruce Harrison and A. Kassianos and P. Sedlak and N. Maugeri and P. J. Goss and J. Davey and David E James and Michael W Graham},
  journal={Methods in enzymology},
  year={2005},
  volume={392},
  pages={
          405-19
        }
}
We describe two complementary strategies for preparing DNA-directed RNA interference (ddRNAi) constructs designed to express hpRNA. The first, oligonucleotide assembly (OA), uses a very simple annealing protocol to combine up to 20 short nucleotides. These are then cloned into appropriately designed restriction sites in expression vectors. OA can be used to prepare simple hairpin (hp)-expressing constructs, but we prefer to use the approach to generate longer constructs. The second strategy… Expand
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References

SHOWING 1-10 OF 15 REFERENCES
Effective expression of small interfering RNA in human cells
TLDR
The simplicity of the U6 expression cassette and its widespread transcription in human cell types suggest that this mode of siRNA delivery could be useful for suppressing expression of a wide range of genes. Expand
A resource for large-scale RNA-interference-based screens in mammals
TLDR
The construction and application of a shRNA expression library targeting 9,610 human and 5,563 mouse genes is reported, which suggests that the large-scale RNAi library can be used in specific, genetic applications in mammals, and will become a valuable resource for gene analysis and discovery. Expand
Retrovirus vector-mediated stable gene silencing in human cell.
TLDR
A retroviral system for delivery of siRNA into cells, which can substantially down-regulate the expression of human p53 gene in human HepG2 cells is described, indicating that retrovirus vector-delivered RNAi may be used in functional genomics and in gene therapy. Expand
A System for Stable Expression of Short Interfering RNAs in Mammalian Cells
TLDR
It is shown that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes. Expand
RNA Interference in Biology and Medicine
First discovered in plants the nematode Caenorhabditis elegans, the production of small interfering RNAs (siRNAs) that bind to and induce the degradation of specific endogenous mRNAs is nowExpand
A large-scale RNAi screen in human cells identifies new components of the p53 pathway
TLDR
The construction of a set of retroviral vectors encoding 23,742 distinct shRNAs, which target 7,914 different human genes for suppression, is reported, which confers resistance to both p53-dependent and p19ARF-dependent proliferation arrest, and abolishes a DNA-damage-induced G1 cell-cycle arrest. Expand
A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA
TLDR
An approach of combining transgenesis by lentiviral vectors expressing siRNAs can be used successfully to generate a large number of mice in which the expression of a specific gene(s) can be down-regulated substantially, and this approach of generating “knockdown” mice will aid in functional genomics. Expand
Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line.
TLDR
Together, the pBabe vectors and omega E cell line should prove useful in experiments where highest frequencies of gene transfer, or concomitant expression of several different genes within a single cell are required with minimal risk of helper virus contamination. Expand
Identification of essential genes in cultured mammalian cells using small interfering RNAs.
TLDR
The first RNAi-induced phenotypes in mammalian cultured cells using RNA interference mediated by duplexes of 21-nt RNAs are reported, and two other lamins, B1 and B2, are now identified as essential proteins. Expand
Approaches for the sequence-specific knockdown of mRNA
TLDR
The major classes of antisense agents currently used by investigators for sequence-specific mRNA knockdowns are antisense oligonucleotides, ribozymes, DNAzymes and RNA interference (RNAi). Expand
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