RNA Interference RNAi | siRNA

the cat sirnas does not eat...? she eats chicken but not her usual dry food. she was about to faint!
RNAi( RNA interference)? When do you think it will be used to start curing diseases such as cancer? If you dont know what RNAi is, check out this link....
What are the uses of RNAi in the different cell types? Can RNAi mechanism protect us from jumping-repetitive elements and from genomic inbalance?
What does RNAi stand for in Biology? Please help me it has something to do with thermodynamics.
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Small GTPase Sar1 is crucial for proglutelin and ?-globulin export from the e...

Small GTPase Sar1 is crucial for proglutelin and ?-globulin export from the endoplasmic reticulum in rice endosperm.

J Exp Bot. 2013 May 16;

Authors: Tian L, Dai LL, Yin ZJ, Fukuda M, Kumamaru T, Dong XB, Xu XP, Qu LQ

Abstract
Rice seed storage proteins glutelin and ?-globulin are synthesized in the endoplasmic reticulum (ER) and deposited in protein storage vacuoles (PSVs). Sar1, a small GTPase, acts as a molecular switch to regulate the assembly of coat protein complex II, which exports secretory protein from the ER to the Golgi apparatus. To reveal the route by which glutelin and ?-globulin exit the ER, four putative Sar1 genes (OsSar1a/b/c/d) were cloned from rice, and transgenic rice were generated with Sar1 overexpressed or suppressed by RNA interference (RNAi) specifically in the endosperm under the control of the rice glutelin promoter. Overexpression or suppression of any OsSar1 did not alter the phenotype. However, simultaneous knockdown of OsSar1a/b/c resulted in floury and shrunken seeds, with an increased level of glutelin precursor and decreased level of the mature ?- and ?-subunit. OsSar1abc RNAi endosperm generated numerous, spherical, novel protein bodies with highly electron-dense matrixes containing both glutelin and ?-globulin. Notably, the novel protein bodies were surrounded by ribosomes, showing that they were derived from the ER. Some of the ER-derived dense protein bodies were attached to a blebbing structure containing prolamin. These results indicated that OsSar1a/b/c play a crucial role in storage proteins exiting from the ER, with functional redundancy in rice endosperm, and glutelin and ?-globulin transported together from the ER to the Golgi apparatus by a pathway mediated by coat protein complex II.

PMID: 23682119 [PubMed - as supplied by publisher]

Selective tracking of template DNA strands after induction of mitosis with unreplicated genomes (MUGs) in Drosophila S2 cells.

Chromosome Res. 2013 May;21(3):329-37

Authors: Drpic D, Barisic M, Pinheiro D, Maiato H

Abstract
According to the "immortal" DNA strand hypothesis (Cairns Nature 255:197-200, 1975), stem cells would keep their template strands in order to prevent the accumulation of mutations, which could occur during DNA replication. Despite the growing number of studies that attempt to test this hypothesis, the conclusions remain highly controversial. In the base of this controversy lie the current limitations of available methodology to selectively and faithfully track the fate of template DNA strands throughout and upon cell division. Here, we developed a method that allows the unequivocal tracking of single chromatids containing template DNA strands in Drosophila S2 cells in culture. This method consists in the induction of mitosis with unreplicated genomes (MUGs) in which cells are allowed to enter mitosis without prior DNA replication. This is achieved by RNAi-mediated knockdown of Double parked, a conserved protein required for the initiation of DNA replication and post-replication checkpoint response. The advantages of this system when compared with MUGs generated in mammalian cells is the preservation of chromatid morphology, the ease of loss-of-function studies and the possibility of in vivo applications. Altogether, this approach allows for the readily visualization and tracking of template DNA strands by simply monitoring cells stably expressing GFP-fusions with either Histone H2B or the centromeric Histone variant CID/CENP-A by time-lapse fluorescence microscopy. This might be useful for the dissection of the molecular mechanism behind asymmetric DNA strand segregation.

PMID: 23681663 [PubMed - in process]

One-step cloning of intron-containing hairpin RNA constructs for RNA interference via isothermal in vitro recombination system.

Planta. 2013 May 17;

Authors: Jiang Y, Xie M, Zhu Q, Ma X, Li X, Liu Y, Zhang Q

Abstract
Hairpin RNA-based RNA interference (hpRNAi) has become a powerful tool for exploring gene function in reverse genetics. Although, several methods are available for making constructs that express hpRNAi, multiple time-consuming cloning steps are usually involved. Here, we introduce an efficient and flexible hpRNAi vector construction method via the isothermal in vitro recombination system (IR-hpRNAi). For an IR-hpRNAi reaction, two PCR products of a target gene sequence are generated, which containS complementary ends (~20 bp) to each other and to the ends of linearized vector, are fused in a way of head-to-head or tail-to-tail into the vector. This IR-hpRNAi method offers two options to construct the RNAi vectors. Using this method, we created a IR-hpRNAi construct for the Arabidopsis PDS3 gene,and verified the silencing effect via Agrobacterium-mediated transformation. The IR-hpRNAi system rules out the requirement of engineering restriction enzyme cutting sites in target DNA fragments, and is ligation-independent. Thus, this method has advantages over the other hpRNAi construction methods.

PMID: 23681019 [PubMed - as supplied by publisher]

Effects of RNA interference-mediated knock-down of hypoxia inducible factor-? on respiratory burst activity of the Pacific oyster Crassostrea gigas hemocytes.

Fish Shellfish Immunol. 2013 May 13;

Authors: Choi SH, Jee BY, Lee SJ, Cho MY, Lee SJ, Kim JW, Jeong HD, Kim KH

Abstract
In mammals, hypoxia-inducible factor-1 ? (HIF-1?) is known to play important roles not only in oxygen homeostasis but also in innate immune responses. In this study, to assess the functional role of HIF-? in respiratory burst activity of Crassostrea gigas hemocytes, oysters were injected with HIF-?- or green fluorescent protein (GFP)-targeted-long double-stranded RNAs (dsRNAs), and at 1, 3, and 7 days post-injection, knock-down of C. gigas HIF-? expression and production of reactive oxygen species (ROS) were analyzed. Expression of HIF-? in mantle, gill, and hemocytes of C. gigas was clearly down-regulated by injection of the HIF-?-targeted-long dsRNA, but was not inhibited by the GFP-targeted-long dsRNA, indicating that HIF-? expression was suppressed through sequence-specific and systemic RNA interference (RNAi). Respiratory burst activity of hemocytes was significantly increased by administration of GFP-targeted-long dsRNA. However, knock-down of HIF-? expression led to significant decrease of chemiluminescence (CL) response of C. gigas hemocytes at 3 and 7 d post-administration of HIF-?-targeted-long dsRNA, indicating the critical role of HIF-? in activation of respiratory burst activity of oyster hemocytes.

PMID: 23680843 [PubMed - as supplied by publisher]

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