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Questions tagged [rna-interference]

A post-transcriptional mechanism of gene-regulation/silencing mediated by small RNAs (siRNA)

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How to map A single N1-methyladenosine (m1A) by primer extension?

I want to map a single N1-methyladenosine (m1A) modification by primer extension. I have silenced a gene which is responsible for guiding modification of m1A then I confirmed the silencing. My next ...
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RNAi termination

Can you stop an ongoing RNAi after transfection with a vector containing siRNA? I want to experiment on gene function loss effect, using RNAi, and gene function gain, using stopping the RNAi.
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How to detect specific mRNA dsRNA by Northern analysis?

I silenced a gene by RNAi method, for that i chose the gene and the size of the gene is 1.5kb and my insert size is 500nts. i did the transfection and i want to confirm the silencing by northern ...
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205 views

Transposons, Viruses and RNA interference

My textbook says that in RNAi, a complementary double stranded RNA sequence attaches to an mRNA and silences it using a protein machinery. I Googled and read about this so now I know what siRNA and ...
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1answer
213 views

Which mRNAs enter P-bodies?

Processing bodies (P-bodies) are distinct foci within the cytoplasm of the eukaryotic cell consisting of many enzymes involved in mRNA turnover (ref). They play a fundamental role in RNA silencing/...
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Do any RNAs directly inhibit transcription

In eukaryotes, microRNAs and small interfering RNAs, as part of protein complexes, can attack specific messenger RNAs with complementary sequences, thereby inhibiting translation. However, RNA can ...
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siRNA designing problem [closed]

I have a gene sequence with 15 alternative splicing forms. I want to design a siRNA for 1 isoform, I select the exon intron junction that is unique for this isoform, and I must design siRNA for this ...
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1answer
141 views

RNA interference and the former Petunias experiment >

I'm having problems trying to understand how RNA interference works. From what I understood, in simple terms there is this dsRNA which is fragmented into siRNA by the enzyme called Dicer, and those ...
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65 views

How much time do the different mechanisms of gene regulation need to take effect?

I am thinking of the major regulatory mechanisms, like general transcription factors, activators, repressors, and RNA interference. If non-active regulator genes using each of the different ...
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56 views

Why do RNAi pathway proteins exist in the nematode when they can cause them harm?

When dsRNA is injected in plant, Dicer cuts it and prevents disease. Why would the plant parasite, M. incognita - a nematode, die when dsRNA was injected in it? It should have saved it. Again, to be ...
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2k views

Small non-coding RNA (sRNA) vs. micro RNA (miRNA)?

What are the differences between micro RNAs (miRNA) and small non-coding RNAs (sRNA)? Are these two terms used interchangeably?
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Which fractions are enriched for siRNA cleavage activity by comparing electrophoresis?

Size exclusion column chromatography was used to separate the proteins in a Drosophila cell lysate to attempt to identify the protein complex responsible for processing long dsRNA into siRNAs. SDS-...
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Are non-coding RNAs introns?

I am slightly confused as to what part of the genome codes for non-coding RNAs. Is it the introns? This would make sense to me as to why they are not transcribed as the introns are not transcribed. Or ...
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167 views

Microinjection method in C. elegans

I have a question about the experiment that using microinjection to inject RNAi into the C. elegans. To prepare the dsRNA for injection, fist have to clone the gene of interest from the worm genome ...
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113 views

RNAi in nematode resistant plants

Background : Certain plants have been genetically engineered to have sense-antisense gene of a parasitic nematode. The dsRNA produced by the plant then inactivates the mRNA produced in the nematode, ...
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84 views

Why introduction of an extra copy of a gene related to pigmentation causes RNA interference in Petunia?

RNAi became famous after Fire and Mello experiment in C.elegans; however, it had been observed before. In the 80's, Jorgensen was trying to increase pigmentation of Petunia flowers by introducing ...
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Can encoding of miRNAs in introns not cause dysregulation of gene expression?

For example, consider a gene X which has an intron which encodes a miRNA that downregulates expression of another gene Y. When the gene X is translated, this miRNA will appear as a byproduct of ...
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965 views

How to Design an siRNA Experiment?

I'm going to undertake an siRNA experiment soon, but I have only read about them. I want to address the role an enzyme plays in processing a protein. From what I understand, I will need to pick two ...
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997 views

Can siRNA induce DNA methylation in mammalian cells?

Some years ago Hiroaki Kawasaki and Kazunari Taira published an article called "Induction of DNA methylation and gene silencing by short interfering RNAs in human cells" in Nature: In plants, ...
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235 views

bi-directional transcription experiment

We suspect a bi-directional transcription event is happening at a locus in our organism where two genes are directly adjacent to each other. The annotation data is not well established. The intergenic ...