shRNAs will also be more specific to whatever you want to silence, depending on how you design them.
Individual miRNAs can have hundreds of different targets that depend on the transcriptional context of a cell and miRNA targeting is still an area of active research. RNA editing can also change either the mature miRNA or the binding site in a target transcript as well.
Emmette
On Thu, Mar 21, 2013 at 12:33 PM, Josiah Zayner <josiah.zayner@gmail.com> wrote:
miRNAs are RNAs so primer synthesis is a no go because that is DNA. You can perhaps reverse transcribe a DNA element and then purify it and then inject/transfect it into a cell but this seems beyond DIYBio at the moment.
miRNAs go through extensive processing in the cell so I think most people tend to use shRNA and other things.
miRNA expression
http://www.origene.com/MicroRNA/
shRNA expression
http://www.genscript.com/siRNA_service.html?gclid=CLv6sZqdjrYCFYFDMgod-lMAEATo view this discussion on the web visit https://groups.google.com/d/msg/diybio/-/MUgdu8PiYYUJ.
On Sunday, March 17, 2013 8:17:16 AM UTC-5, Mega wrote:Hi,--
although I don't have any projects at the moment in this direction, I'm quite interested in miRNAs...
Can one design a mRNA like this:
If you have a promoter, there's the transcription initiation site where the promoter ends. Next to it you wouldn't attach any ribosome binding sites (because all you are interested in is the RNA, protein synthesis would be waste of energy). Then I think you would add the complementary sequence of the CDS (the first 20 nucleotides or so), excluding the ATG so it won't have a chance to silence other proteins.
Should work like this, right?
What makes it specifically interesting is that they are so short you basically could have them synthesized as primers, in this size without expensive purification...
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