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Not RNAi. Has anyone tried gene knock-up? - (Feb/11/2013 )

We have some genes that we want to test at higher levels of expression. Apart from knock-in, does anyone have any idea how to do this? SINEUP would be one way, but its quite a new technology so I was wondering if anyone has given it a try?


There is a new technique applicable for some genes -- inhibiting natural antisense transcripts to change regulation of their complementary mRNAs.

Inhibition of natural antisense transcripts in vivo results in gene-specific transcriptional upregulation.

Modarresi F, Faghihi MA, Lopez-Toledano MA, Fatemi RP, Magistri M, Brothers SP, van der Brug MP, Wahlestedt C.
Nat Biotechnol. 2012 Mar 25;30(5):453-9. doi: 10.1038/nbt.2158.

Another more established possibility, if the mRNA of interest is under miRNA repression, is to protect the site on the mRNA where the miRNA binds using an oligo as a target protector.

Target protectors reveal dampening and balancing of Nodal agonist and antagonist by miR-430.

Choi WY, Giraldez AJ, Schier AF. Science. 2007 Oct 12;318(5848):271-4. Epub 2007 Aug 30.

-Jon Moulton-

Try an inducible promoter on an inserted plasmid.