Lentivial Knockdown of miRNAs
Posted 07 January 2010 - 01:28 AM
Also, do you know what should be cloned into the miRZip vector? Thanks.
Posted 07 January 2010 - 11:36 AM
Posted 07 January 2010 - 05:58 PM
Two options are to make an shRNA targeting the loop section of the pre-miRNA molecule, or clone in a synthetic miRNA target for the miRNA to bind to (i.e. like a microRNA sponge). I'm sure that there are other possibilities though...
THanks for your reply, miRNA man. Could you explain a bit more on that, or are there any references/links that I can take a look?
Posted 08 January 2010 - 11:33 AM
Posted 30 March 2010 - 08:52 PM
Posted 01 April 2010 - 06:43 PM
Posted 01 April 2010 - 09:21 PM
Thank you,pcrman!I am constructing a spnoges for a miRNA , only promoter + sponges without luciferase, and hope it could work well!
Your can read the original paper from Philp Sharp lab. You need to clone multiple copies of the miRNA target into the 3UTR of a gene, for example, luciferase.
Posted 02 April 2010 - 01:48 PM
Posted 02 April 2010 - 05:21 PM
Thank you! At first, I think I could detect the amount of mature miRNA to test the sponges work, Now I will add a report gene on upstream of Sponges.
Why don't you want to include a reporter gene like luc as pcrman suggested? It should be useful to track expression and besides, I personally doubt if a sponge would work without a gene (the mRNA needs to be exported to the cytoplasm to be functional...will this happen if there is no CDS?)
To miR ,I am a new. Thank you!