I thougth of this:
Promoter- UTR-miRNA- Terminator.
RNA-Polymerase is "loading" at the promoter, and then it starts the translation. Some part of the MCS is also transcribed. Then there is the reverse complement of the DNA, so this gets transcribed into RNA. then there is still some UTR and then some polyA.
So basically you get AGTGCTGCT(junk) - ATAGTCGCATCG (miRNA) - TCGTCGTC (junk) The piece in the middle is complementary to the long protein coding RNA, and binds to it.
On Thu, Mar 21, 2013 at 5:33 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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